Design and synthesis of 5-chloro-2-hydroxy-3-triazolylbenzoic acids as HIV integrase inhibitors

Article abstract of DOI:10.1007/s00044-015-1325-9

A series of potential HIV-1 integrase inhibitors based on 5-chloro-2-hydroxy-3-triazolylbenzoic acid scaffold was designed and synthesized. Some of these compounds exhibit potent inhibitory activities at micromolar concentrations against HIV-1 integrase i

Full text of DOI:10.1007/s00044-015-1325-9

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res
DOI 10.1007/s00044-015-1325-9
ORIGINAL RESEARCH
Design and synthesis of 5-chloro-2-hydroxy-3-triazolylbenzoic
acids as HIV integrase inhibitors
Jie Chen
Wei Liu
•
Cheng-Fu Liu
Li-Ming Hu
•
Cheng-Wen Yang
•
Cheng-Chu Zeng
•
•
Received: 23 September 2014 / Accepted: 16 January 2015
Ó Springer Science+Business Media New York 2015
Abstract A series of potential HIV-1 integrase inhibitors
based on 5-chloro-2-hydroxy-3-triazolylbenzoic acid scaf-
fold was designed and synthesized. Some of these com-
pounds exhibit potent inhibitory activities at micromolar
2004); therefore, it is essential for HIV-1 replication. More-
over, since there is no corresponding known human counter-
part, HIV IN is regarded as a promising chemotherapeutical
target to treat AIDS (Esposito and Craigie, 1999; Gupta and
Nagappa, 2003; Anthony, 2004).
0
concentrations against HIV-1 integrase in the 3 -end pro-
cessing and the strand transfer step.
During the past decades, much attention has been paid to
the design and synthesis of HIV IN inhibitors and a large
number of compounds, with great structural diversity, have
been synthesized and evaluated as possible HIV IN in-
hibitors, which lead to the discovery of many typical HIV
IN inhibitors, including aryl b-diketo acids (such as s-1360
and L-708,906) (Fig. 1) (Hazuda et al., 2004; Bacchi et al.,
Graphical Abstract
Cl
IC : 28.00 uM (L-708,906)
R
50
IC : 1.56 uM (5f)
50
HOOC
N
N
IC50: 3.50 uM (5v)
N
OH
IC : 5.70 uM (5aa)
50
2
011a, b), styrylquinones (Fig. 1) (Mouscadet and Des-
maele, 2010), polyhydroxylated aromatics (Wang et al.,
009; Yang et al., 2010; Reddy et al., 1999) and
5a-5ad
2
oligonucleotides (Drake et al., 1998; Jing et al., 2000).
Among these discoveries and advances, the most repre-
sentative achievement is the disclosure of the keto-enol
acid scaffold for the IN inhibitory activity, and the most
successful one is the raltegravir (Fig. 1), the first integrase
inhibitor (INI) approved by the Food and Drug Adminis-
tration (FDA) for the treatment of AIDS (Evering and
Markowitz, 2008; Temesgen and Siraj, 2008).
Keywords Click chemistry ꢀ Diketo acids ꢀ
HIV IN inhibitors ꢀ 2-Hydroxy-3-triazolylbenzoic acids ꢀ
Metal ion binding
Introduction
The general structure of aryl b-diketo acid inhibitors,
taking L-708,906 as an example, is comprised of a phar-
macophore(s) and an appending hydrophilic sub-
unit(s) (Fig. 2, top) (Kawasuji et al., 2006a, b). An indole
ring or a benzene ring appended by one or two arylalkyl
substituents is a typical hydrophobic moiety, which has a
specific interaction with an adjacent hydrophobic pocket or
surface. Meanwhile, the pharmacophore, typically being
aryl diketo acid or its bioisoster, functions through a
competitive binding of two divalent metallic ion in the IN
active core, and thus blocking the access of a host DNA to
the integrase enzyme.
HIV-1 integrase (IN), along with reverse transcriptase and
protease, are encoded by HIV genome. HIV IN functions
through catalyzing the insertion and integration of the proviral
0
DNA into the genome of the host cell in 3 -end processing and
strand transfer steps (LaFemina et al., 1992; Chiu and Davies,
LJ .. -CMh .e nH uꢀ C.-F. Liu ꢀ C.-W. Yang ꢀ C.-C. Zeng (&) ꢀ W. Liu ꢀ
College of Life-Sciences and Bioengineering, Beijing University
of Technology, Beijing 100124, China
e-mail: zengcc@bjut.edu.cn; zengcc138@gmail.com
123

Med Chem Res
Fig. 1 The structures of some
typical HIV IN inhibitors
F
O
O
OH
O
O
N
NH
N
O
COOH
HO
S1360
L708906
N
O
N
OH
F
OH
OH
N
N
HOOC
N
H
N
H
OH
N
O
O
O
styrylquinone
raltegravir
Fig. 2 Design of 5-chloro-2-
hydroxy-3-triazolylbenzoic
acids as HIV integrase
inhibitors
O
O
Hydrophilic subunit
(pharmacophore)
Hydrophobic subunit
O
O
O
O
L-708,906
Responsible for the quick construction
of two subunit through "Click Reaction"
H
N
Hydrophilic subunit
pharmacophore)
(
Hydrophobic subunit
O
O
R
N
N
N
O-
O
N
I
Hydrophobic subunit
Increasing the acidic
properties of the phenolic OH
Cl
O
N
N
N
Pharmocophore with two-
metal chelation sites
O
O
II
Responsible for the quick
construction of two subunit and
participation of chelating metal ion
Based on the understanding of the structural feature of
diketo acid type of HIV IN inhibitors, we have carried out a
project (Xu et al., 2009; Zeng et al., 2009) with the basic
assumption that the development of HIV IN inhibitors can
be in principle performed by a combination of appropriate
pharmacophore and a hydrophobic moiety linked by an
efficient method, such as click chemistry (Hou et al., 2012;
Bock et al., 2005). In this regard, we have designed and
expected anti-HIV IN activity was not observed (Fig. 2,
middle) (Zeng et al., 2009).
Continuing on the basis of the previous project, we
further modified structure I, the pharmacophore, to struc-
ture II, a 5-chloro-2-hydroxy-3-triazolylbenzoic acid sub-
unit. In this design, the N-5 position of the former is
replaced by an aromatic carbon atom and the fused pyr-
idine ring is modified to be a triazolyl group (Fig. 2, bot-
tom). We anticipate that the triazole ring plays a double
role, namely, it does not only serve as a linker but would
also participate in metal ion binding. In addition, to
synthesized
a
series of 1,4-dihydro-4-oxo-1,5-naph-
thyridine-3-carboxylic acid derivatives (see structure I in
Fig. 1, middle) as potential HIV IN inhibitors; however,
1
23

Med Chem Res
increase the acidic properties of the phenolic OH at C-2
position, a chloride atom was introduced on the C-5 of the
benzoic acid (Jiao et al., 2010). Herein, we reported the
synthesis of 5-chloro-2-hydroxy-3-triazolylbenzoic acids.
The preliminary HIV IN inhibitory activity evaluation
shows that some of these compounds can inhibit the
replication of HIV-1 in vitro against integrase at micro-
molar concentration (Zeng et al., 2010).
and compounds 5k–5r benzyloxymethyl groups, whereas, in
the cases of 5t–5ad, a dibenzylaminomethyl group is intro-
duced (benzylaminomethyl for 5s).
All title compounds 5a–5ad were preliminarily tested
against purified HIV IN to determine any possible inhibitory
activity, and the results are summarized in Table 1. Under
the activity evaluation conditions, the IC50 values of com-
pounds L-708,906 and raltegravir are 28.0 and 0.00427 lg/
mL, respectively, notably, the IC50 of L-708,906 for strand
transfer is *0.06 lM (Marchand et al., 2003). It was ob-
served that the HIV IN inhibitory effect of compound 5a and
most of the O-bridged compounds (5b–5d and 5g–5r) were
less than 50 % at an initial concentration of 20 lg/mL.
Structurally, these O-bridged compounds possess only one
side-chain (whereas two side-chain for L-708,906), which
may lead to lower hydrophobic interaction with the hy-
drophobic pocket of HIV IN compared with the positive
compound. To our delight, most of the N-bridged compounds
(5t–5v and 5w–5ad) exhibit a higher potency than
L-708,906. For example, the IC50 of compound 5z (bearing
two o-fluorobenzyl groups) and 5ac (bearing two
o-bromobenzyl groups) are 17.5 lM and 12.8 lM, respec-
tively. In addition, the corresponding 5p (bearing one
o-fluorobenzyl group) and 5r (bearing one o-bromobenzyl
group) show less than 50 % inhibitory effect at initial 20 lg/
mL. As expected, this outcome may be the result of the in-
crease in hydrophobic interaction of the N-bridged com-
pounds with the hydrophobic pocket of HIV IN.
Results and discussion
The synthesis of 5-chloro-2-hydroxy-3-triazolylbenzoic
acids 5 is depicted in Scheme 1. Nitration of 5-chloro-2-hy-
droxybenzoicacid1gave5-chloro-2-hydroxy-3-nitrobenzoic
acid2, which was reducedbyNa S O atroom temperature to
2
2 4
affordcompound3. The keyintermediate azidobenzoicacid4
was produced in a good yield by using tert-butyl nitrite and
azidotrimethylsilane under mild conditions (Barral et al.,
2
007). Target 5-chloro-2-hydroxy-3-triazolylbenzoic acids
5
a–5ad were finally obtained in good to excellent yields by
copper (I)-catalyzed Huisgen 1,3-dipolar cycloaddition of 4
and terminal alkynes (Scheme 1; Table 1).
As mentioned above, we selected 5-chloro-2-hydroxy-3-
triazolylbenzoic acid subunit as the pharmacophore wherein
the triazole ring may function not only as part of the phar-
macophore, but also as a linker to assemble the pharma-
cophore and hydrophobic unit. The hydrophobic subunit was
the main modified moiety, and as a result, the designed target
compounds 5a–5ad differ only in the structure of the side
chain attached to the triazolyl ring (both as a whole using as
hydrophobic subunit). As shown in Table 1, for compound
In addition, the screening results reveal that the IC50 of
compounds 5v and 5aa are less than 6.0 lM, and the best
result is afforded for p-bromophenoxy methyl triazolyl-
benzoic acid (5f), with an IC50 of 1.56 lM.
5
a, a benzene ring appends directly to the triazole unit. In
order to adjust its flexibility and improve the possible hy-
drophobic interaction with the hydrophobic pocket of HIV
IN, the side chain of the target compounds is introduced
through oxygen or nitrogen atom to the triazole ring. For
example, 5b–5j are substituted by phenoxymethyl groups
Conclusion
In summary, HIV IN inhibitors based on 5-chloro-2-hy-
droxy-3-triazolylbenzoic acid scaffold were designed; their
Scheme 1 Synthesis of
-chloro-2-hydroxy-3-
triazolylbenzoic acids 5a–5ad
Cl
Cl
Cl
5
HNO /H SO
3
2
4
Na S O
2 2 4
HOOC
rt
HOOC
(
NO2
CH OH/H O
HOOC
NH2
3
2
OH
1
OH
2
OH
3
(Yield: 38%)
Yield: 96%)
Cl
Cl
R1
t-BuONO, TMSN3
terminal alkyne
HOOC
N
N
HOOC
N3
CH CN
sodium ascorbate, CuSO4
N
3
OH
OH
4
t-BuOH/H O
2
5a-5ad
(Yields: 45-90%)
(Yield: 88%)
123

Med Chem Res
Table 1 Inhibitory activity results of target compounds 5a–5ad in vitro
Structure of compound
IC50
Structure of compound
raltegravir
IC50
(
μg/mL)
(μg/mL)
0.00427
O
O
O
28.0
COOH
O
L-708906
O
Phar
Phar
–
N
N
N
–
F
N
N
N
N
N
5
a
5p
N
N
OH
F
O
Phar
N
5b
–
–
Phar
–
N
5q
Br
O
O
Phar
N
N
N
Phar
Phar
–
5c
N
N
N
5r
NH
O
Phar
Phar
N
N
N
N
N
N
2
9.1
1.6
.56
33.1
26.3
18.4
5
d
5s
O
N
N
N
Phar
N
N
N
2
1
5e
N
t-Bu
5
t
O
CH
3
Phar
N
N
N
N
5
Phar
f
N
N
Br
N
CH
3
5u
O
Phar
N
N
N
N
Me
Me
–
–
Phar
3.50
23.7
5g
N
N
NO
2
N
5
v
Me
O
Phar
N
N
N
N
Phar
O
N
N
5
h
N
Me
5w
F
O
O
N
Phar
N
N
–
Phar
19.0
N
N
N
5
i
N
5
x
F
1
23

Med Chem Res
Table 1 continued
O
O
N
Phar
F
F
N
N
–
–
Phar
38.2
17.5
N
N
N
5j
N
5
y
F
O
N
Phar
N
N
Phar
5k
N
N
N
N
5z
F
O
N
NO
2
Phar
Me
N
N
Phar
N
N
32.9
–
N
5.70
13.8
5
l
N
N
NO
2
5aa
O
N
Br
Phar
Phar
N
Me
N
N
N
5m
N
Br
5
ab
Br
Me
O
N
34.8
Phar
12.8
19.8
N
N
Phar
Phar
N
N
N
N
5n
N
5ac
Br
Cl
O
N
N
F
49.3
Phar
N
5o
N
N
N
5
ad Cl
For clarity, Phar in the Table is referred to 5-chloro-2-hydroxy-benzoic acid
‘‘–’’ The HIV-IN inhibitory effect was less than 50 % at the initial concentration of 20 lg/mL
synthesis follows a straightforward pathway by using a
click chemistry reaction. Compared with O-bridged com-
pounds, most of the N-bridged compounds inhibit replica-
tion of HIV -1 integrase more efficiently than the positive
compound L-708,906. The work has demonstrated that our
click chemistry reaction methodology has been quite useful
in the design of HIV IN inhibitors, producing a promising
new starting point for further optimization.
recorded on an AV 400M Bruker spectrometer (400 MHz for
1
13
H frequency, 100 MHz for C frequency) in solvent
(CDCl , CD OD or DMSO-d ) with TMS as an internal
3
3
6
reference. ESI–MS was measured on a Bruker esquire 6000
mass spectrometer. All solvents were available from com-
mercial source and used directly without further purification.
All starting terminal alkynes (Trost and Xie, 2006; Inamoto
et al., 2005; Dinges et al., 2007; Ma et al., 2005; Brouillette
et al., 1994) and compounds 2 (Musser et al., 1985) and 3
(Von Plessing and Carlos, 1963) were synthesized according
Experimental
to known procedures. Cautions for using TMSN : to work in
3
a fume hood with good ventilation.
Materials and methods
Synthesis of 3-azido-5-chloro-2-hydroxybenzoic acid
All melting points were measured using an XT4A elec-
trothermal instrument equipped with a microscope and are
uncorrected. Infrared spectra (IR) were measured as thin
films on KBr plates on a Bruker IR spectrophotometer and
(4)[31]
In a 100-mL round-bottom flask was added 50 mL of
acetonitrile containing 3-amino-5-chloro-2-hydroxyben-
zoic acid (0.94 g, 5 mmol) and cooled in an ice-water bath.
-
1
1
13
are expressed in v (cm ). H and C NMR spectra were
123

Med Chem Res
To the flask was added dropwise t-BuONO (7.5 mmol,
75 lL) and TMSN (6 mmol, 755 lL). After stirring for
J = 2.8 Hz, Ar–H), 8.01 (d, 1H, J = 2.8 Hz, Ar–H), 8.69
1
3
9
(s, 1H, triazole-H); C NMR (100 MHz, DMSO-d ): d
6
3
about 30 min, the ice-water bath was removed and the
stirring was continued for about 3 h. Upon completion,
most of the solvents were evaporated under reduced pres-
sure and the product then was precipitated to give gray
solid. The reaction mixture was filtered and washed with
petroleum ether and then water. After drying under
61.2, 115.2, 121.4, 122.4, 126.8, 130.0, 130.3, 130.5,
143.4, 153.5, 158.5, 170.8; IR (KBr): m 3,438, 2,927, 1,638,
1,472 cm ; MS (ESI): m/z 343.6 [M-H] .
-
1
-
5
1
2
2
-Chloro-2-hydroxy-3-[4-(naphthalen-1-yloxymethyl-1H-
,2,3-triazol-1-yl)benzoic acid (5d) Yield: 65 %; M. P.:
1
31–233 °C; H NMR (400 MHz, DMSO-d ): d 5.44 (s,
6
vacuum, a red solid (938 mg) was obtained in 88 % yield.
1
H, OCH ), 7.22–7.53 (m, 4H, Ar–H), 7.86 (d, 1H,
2
M.P.: 138–140 °C; H NMR (400 MHz, DMSO-d ): d 7.35
6
J = 7.6 Hz, Ar–H), 7.91 (d, 1H, J = 2.4 Hz, Ar–H), 8.04
(
d, 1H, J = 2.8 Hz, Ar–H), 7.54 (d, 1H, J = 2.8 Hz, Ar–
(
d, 1H, J = 2.4 Hz, Ar–H), 8.13 (d, 1H, J = 8.4 Hz, Ar–
1
H), 12.41 (bs, 1H, COOH); C NMR (100 MHz, DMSO-
3
1
3
H), 8.82 (s, 1H, triazole-H); C NMR (100 MHz, DMSO-
d6): d 115.6, 122.9, 125.8, 125.9, 129.3, 153.7, 171.2; IR
d6): d 62.0, 106.4, 120.9, 122.0, 122.2, 125.4, 125.8, 126.6,
(
KBr): m 3,439.6, 2,135, 1,676, 1,464, 1,292, 1,232,
1
1
26.7, 126.8, 126.9, 127.9, 130.2, 130.5, 134.5, 143.4,
1
-
,171 cm ; MS (ESI): m/z 211.6 [M-H] .
1
-
1
-
53.8, 154.0, 170.8; IR (KBr): m 3,437, 1,634 cm ; MS
-
ESI): m/z 393.7 [M-H] ; HRMS (ESI) calcd for
(
C H ClN O [M?H]: 396.0751, found: 396.0750.
2
0
15
3 4
The general procedure for the synthesis of 5-chloro-2-
hydroxy-3-triazolylbenzoic acids (5)
5-Chloro-2-hydroxy-3-[4-(4-tert-butylphenoxymethyl)-1H-
,2,3-triazol-1-yl]benzoic acid (5e) Yield: 48 %; M. P.:
1
1
In a 250-mL three-necked round-bottom flask was added
226–228 °C; H NMR (400 MHz, DMSO-d
6
): d 1.24 (s,
9H, C(CH ), 5.19 (s, 2H, OCH ), 6.98 (d, 2H,
2
0 mL of tert-butanol/water (V:V = 1:1) mixed solution
containing 3-azido-5-chloro-2-hydroxybenzoic acid
427 mg, 2 mmol) and terminal alkyne (2.2 mmol). To this
solution were then added sodium ascorbate (200 mg,
3
)
3
2
J = 8.8 Hz, Ar–H), 7.29 (d, 2H, J = 8.8 Hz, Ar–H), 7.83
(d, 1H, J = 2.0 Hz, Ar–H), 7.89 (d, 1H, J = 2.0 Hz, Ar–
(
1
3
H), 8.74 (s, 1H, triazole-H); C NMR (100 MHz, DMSO-
): d 31.8, 34.2, 61.4, 114.7, 120.2, 126.3, 126.6, 126.7,
128.4, 130.1, 143.3, 143.5, 155.4, 156.3, 170.8; IR (KBr): m
1
0
mmol, 50 % equiv.) and CuSO ꢀ5H O (50 mg,
d
6
4
2
.2 mmol, 10 % equiv.). The reaction mixture was stirred
-
1
-
for about 8–12 h at room temperature. The precipitate was
filtered, washed with water and then petroleum ether, and
dried under vacuum to give gray solid.
3,436, 1,633 cm ; MS (ESI): m/z 399.7 [M-H] .
5-Chloro-2-hydroxy-3-[4-(4-bromophenoxymethyl)-1H-1,2,3-
triazol-1-yl]benzoic acid (5f) Yield: 90 %; M. P.:
1
5
-Chloro-2-hydroxy-3-(4-phenyl-1H-1,2,3-triazol-1-yl)benzoic
1
2
34–236 °C; H NMR (400 MHz, MeOH-d ): d 5.19 (s, 2H,
4
acid (5a) Yield: 69 %; M.P.: 260–262 °C; H NMR
400 MHz, DMSO-d ): d 7.36 (t, 1H, J = 7.2 Hz, Ar–H),
OCH ), 6.95 (d, 2H, J = 8.8 Hz, Ar–H), 7.38 (d, 2H,
2
(
6
J = 8.4 Hz, Ar–H), 7.83(s, 1H, Ar–H), 8.00 (s, 1H, Ar–H), 8.57
13
7
.47 (t, 2H, J = 7.6 Hz, Ar–H), 7.77 (d, 2H, Ar–H), 7.95
1
(
s, 1H, triazole-H); C NMR (100 MHz, DMSO-d ): d 61.3,
3
6
(
d, 2H, J = 7.6 Hz, Ar–H), 9.12 (s, 1H, triazole-H);
C
1
1
1
8
15.3, 117.5, 121.2, 124.2, 125.6, 125.7, 126.7, 129.2, 130.5,
NMR (100 MHz, DMSO-d ): d 117.5, 121.1, 124.3, 125.5,
6
32.2, 148.6, 156.3, 170.1; IR (KBr): m 3,437, 2,925, 1,639,
,488, 1,469, 1,424, 1,384, 1,241, 1,173, 1,102, 1,074, 1,024,
1
1
25.9, 126.8, 128.6, 129.2, 130.8, 132.1, 148.4, 157.6,
-
69.5; IR (KBr): m 3,437, 1,637, 1,469, 1,418, cm ; MS
1
-1
86, 817, 776, 727, 527 cm ; MS (ESI): m/z 421.6 [M-H] .
-
-
(
ESI): m/z 313.6 [M-H] .
5-Chloro-2-hydroxy-3-[4-(4-nitrophenoxymethyl)-1H-1,2,3-
5
-Chloro-2-hydroxy-3-[4-(hydroxymethyl)-1H-1,2,3-triazol-1-
1
triazol-1-yl]benzoic acid (5g) Yield: 71 %: M. P.:
1
yl]benzoic acid (5b) Yield: 52 %; M.P.: 175–177 °C; H
NMR (400 MHz, DMSO-d ): d 4.60 (s, 2H, OCH ),
[300 °C; H NMR (400 MHz, DMSO-d ): d 5.41 (s, 2H,
6
6
2
OCH ), 7.31 (d, 2H, J = 8.4 Hz, Ar–H), 7.75 (s, 2H, Ar–
2
1
3
7
.70–7.73 (m, 2H, Ar–H), 8.58 (s, 1H, triazole-H);
C
H), 8.23 (d, 2H, J = 8.8 Hz, Ar–H), 8.90 (s, 1H, triazole-
1
3
NMR (100 MHz, DMSO-d ): d 55.4, 117.7, 122.2, 124.2,
H); C NMR (100 MHz, DMSO-d ): d 62.3, 115.8, 118.0,
6
6
1
25.8, 126.6, 129.4, 148.1, 157.3, 169.7; IR (KBr): m 3,435,
,635 cm ; MS (ESI): m/z 267.5 [M-H] .
126.3, 126.4, 126.6, 129.9, 141.5, 141.9, 157.3, 163.8,
-1
-
1
-
1
169.7; IR (KBr): m 3,438, 1,637 cm ; MS (ESI): m/z
-
88.6 [M-H] ; HRMS (ESI) calcd for C H ClN O
3
1
6
12
4
6
5
-Chloro-2-hydroxy-3-[4-(phenoxymethyl)-1H-1,2,3-triazol-
[M?H]: 391.0445, found: 396.0509.
1
-yl]benzoic acid (5c) Yield: 69 %; M. P.: 230–232 °C;
1
H NMR (400 MHz, DMSO-d ): d 5.23 (s, 2H, OCH ),
5-Chloro-2-hydroxy-3-{4-[(4-prop-2-ynyloxy)phenoxy]methyl-
6
2
6
.95 (t, 1H, J = 7.2 Hz, Ar–H), 7.07 (d, 2H, J = 8.0 Hz,
1H-1,2,3-triazol-1-yl}benzoic acid (5h) Yield: 42 %; M. P.:
1
Ar–H), 7.30 (t, 2H, J = 8.0 Hz, Ar–H), 7.91 (d, 1H,
217–219 °C; H NMR (400 MHz, DMSO-d ): d 3.51 (t,
6
1
23

Med Chem Res
1
H, J = 2.4 Hz, : C–H), 4.72 (d, 2H, J = 2.4 Hz, : C–
CH ), 5.17 (s, 2H, OCH ), 6.94 (d, 2H, J = 8.8 Hz, Ar–H),
5-Chloro-2-hydroxy-3-{4-[(3-methylbenzyloxy)methyl]-1H-
1,2,3-triazol-1-yl}benzoic acid (5m) Yield: 69 %; M. P.:
1
2
2
7
.02 (d, 2H, J = 8.8 Hz, Ar–H), 7.76 (s, 2H, Ar–H), 8.80
3
190–192 °C; H NMR (400 MHz, DMSO-d ): d 2.30 (s,
6
1
(
s, 1H, triazole-H); C NMR (100 MHz, DMSO-d ): d
3H, Ar–CH ), 4.55 (s, 2H, OCH Ar), 4.67 (s, 2H, OCH ),
6
3
2
2
5
6.4, 61.9, 78.4, 80.0, 116.1, 116.4, 117.6, 122.1, 126.0,
7.10–7.17 (m, 3H, Ar–H), 7.25 (t, 1H, J = 7.2 Hz, Ar–H),
1
26.1, 126.5, 129.7, 143.0, 152.0, 153.1, 157.6, 169.6; IR
-
7.86 (s, 1H, Ar–H), 7.93 (s, 1H, Ar–H), 8.64 (s, 1H, Ar–H);
13
1
(KBr):m 3,436, 1,637, 1,507 cm ; MS (ESI): m/z 397.6
[M-H] .
C NMR (100 MHz, DMSO-d ): d 21.4, 63.2, 71.8, 120.7,
6
-
125.2, 126.0, 128.6, 128.6, 128.7, 128.9, 130.1, 137.8,
1
1
38.5, 155.0, 170.2; IR (KBr): m 3,436, 1,637, 1,472,
,423 cm ; MS (ESI): m/z 371.6 [M-H] ; HRMS (ESI)
5
1
2
1
-Chloro-2-hydroxy-3-{4-[(3-prop-2-ynyloxy)phenoxy]methyl-
-
1
-
H-1,2,3-triazol-1-yl}benzoic acid (5i) Yield: 80 %; M. P.:
calcd for C H ClN O [M?H]: 374.0908, found:
18 17 3 4
1
10–211 °C; H NMR (400 MHz, DMSO-d ): d 3.53 (t,
6
374.0894.
H, J = 2.4 Hz, : C–H), 4.78 (d, 2H, J = 2.4 Hz, : C–
CH ), 5.22 (s, 2H, OCH ), 6.59–6.61 (m, 1H, Ar–H), 6.70
2
5-Chloro-2-hydroxy-3-{4-[(2-methylbenzyloxy)methyl]-1H-
2
(
s, 1H, Ar–H), 6.71–6.72 (m, 1H, Ar–H), 7.22 (t, 1H,
1,2,3-triazol-1-yl}benzoic acid (5n) Yield: 54 %; M. P.:
1
J = 8.4 Hz, Ar–H), 7.76 (d, 1H, J = 2.8 Hz, Ar–H), 7.78
1
(
195–197 °C; H NMR (400 MHz, DMSO-d6): d 2.27 (s,
3
d, 1H, J = 2.8 Hz, Ar–H), 8.82 (s, 1H, triazole-H);
C
3H, Ar–CH ), 4.58 (s, 2H, OCH Ar), 4.68 (s, 2H, OCH ),
3
2
2
NMR (100 MHz, DMSO-d ): d 56.0, 61.6, 78.7, 79.8,
7.18–7.34 (m, 4H, Ar–H), 7.86 (d, 1H, J = 2.4 Hz, Ar–H),
7.93 (d, 1H, J = 2.4 Hz, Ar–H),8.63 (s, 1H, triazole-H);
6
1
1
1
02.5, 108.1, 117.8, 122.2, 126.2, 126.3, 126.5, 129.8,
1
3
30.5, 142.8, 157.5, 159.0, 159.8, 169.7; IR (KBr): m 3,437,
,636, 1,469 cm ; MS (ESI): m/z 397.6 [M-H] .
C NMR (100 MHz, DMSO-d ): d 18.8, 63.2, 70.3, 121.2,
6
-
1
-
125.9, 126.0, 126.7, 128.1, 128.9, 129.0, 130.2, 130.4,
1
36.5, 136.8, 144.5, 154.5, 170.5; IR (KBr): m 3,437, 1,637,
5
1
2
1
-Chloro-2-hydroxy-3-{4-[(2-prop-2-ynyloxy)phenoxy]methyl-
-
,471, 1,422 cm ; MS (ESI): m/z 371.8 [M-H] ; HRMS
1
-
1
H-1,2,3-triazol-1-yl}benzoic acid (5j) Yield: 56 %; M. P.:
1
(ESI) calcd for C H ClN O [M?H]: 374.0908, found:
18 17 3 4
06–208 °C; H NMR (400 MHz, DMSO-d ): d 3.50 (t,
6
374.0886.
H, J = 2.4 Hz, : C–H), 4.78 (d, 2H, J = 2.4 Hz, : C–
CH ), 5.23 (s, 2H, OCH ), 6.93–7.24 (m, 4H, Ar–H), 7.78
2
5-Chloro-2-hydroxy-3-{4-[(4-fluorobenzyloxy)methyl]-1H-
2
1
3
(
s, 2H, Ar–H), 8.82 (s, 1H, triazole-H);
C NMR
1,2,3-triazol-1-yl}benzoic acid (5o) Yield: 51 %; M. P.:
1
(
100 MHz, DMSO-d ): d 56.6, 62.1, 78.6, 79.9, 114.8,
201–203 °C; H NMR (400 MHz, CD OD): d 4.60 (s, 2H,
6
3
1
1
1
15.3, 117.9, 121.6, 121.9, 122.3, 126.2, 126.3, 126.5,
OCH Ar), 4.72 (s, 2H, OCH ), 7.05–7.09 (m, 2H, Ar–H),
2
2
29.8, 142.9, 147.4, 148.6, 157.4, 169.7; IR (KBr): m 3,437,
,637, 1,502 cm ; MS (ESI): m/z 397.6 [M-H] ;
7.38–7.41 (m, 2H, Ar–H), 7.85 (s, 1H, Ar–H), 8.02 (s, 1H,
13
Ar–H), 8.52 (s, 1H, triazole-H); C NMR (100 MHz,
-
1
-
DMSO-d ): d 63.2, 71.0, 115.4, 115.6, 120.0, 126.3, 127.0,
6
5
-Chloro-2-hydroxy-3-[4-(benzyloxymethyl-1H-1,2,3-triazol-
1
28.0, 130.2, 130.3, 134.8, 134.8, 155.4, 160.8, 163.3,
1
1
-yl)]benzoic acid (5k) Yield: 85 %; M. P.: 225–227 °C;
-
1
70.3; IR (KBr): m 3,437, 1,638, 1,511,1,471 cm ; MS
1
H NMR (400 MHz, DMSO-d ): d 4.59 (s, 2H, OCH Ar),
6
2
-
ESI): m/z 375.9 [M-H] .
(
4
.69 (s, 2H, OCH ), 7.31–7.38 (m, 5H, Ar–H), 7.93–7.99
2
1
m, 2H, Ar–H), 8.68 (s, 1H, triazole-H); C NMR
3
(
5-Chloro-2-hydroxy-3-{4-[(3-fluorobenzyloxy)methyl]-1H-
(
100 MHz, DMSO-d ): d 63.2, 71.8, 127.9, 127.9, 128.1,
1,2,3-triazol-1-yl}benzoic acid (5p) Yield: 67 %; M.P.:
1
6
1
28.6, 128.7, 138.5, 169.7; IR (KBr): m 3,436, 1,638,
,471 cm ; MS (ESI): m/z 357.8 [M-H] , 359.8
204–206 °C; H NMR (400 MHz, DMSO-d6): d 4.61 (s,
-
1
-
1
2H, OCH Ar), 4.70 (s, 2H, OCH ), 7.12–7.41 (m, 4H, Ar–
2
2
?
M?H] , HRMS: m/z calcd for C H N O Cl:
[
H), 7.92 (s, 1H, Ar–H), 8.02 (s, 1H, Ar–H), 8.63 (s, 1H,
1
1
7 14 3 4
3
3
58.06001; found: 358.0590.
triazole-H); C NMR (100 MHz, DMSO-d ): d 63.1, 71.2,
6
1
1
1
15.3, 115.7, 120.2, 126.1, 127.0, 128.2, 130.1, 130.3,
5
1
1
3
7
-Chloro-2-hydroxy-3-{4-[(4-methylbenzyloxy)methyl]-1H-
34.7, 134.8, 155.3, 160.9, 163.1, 170.0; IR (KBr): m 3,437,
,638, 1,471 cm ; MS (ESI): m/z 375.8 [M-H] .
,2,3-triazol-1-yl}benzoic acid (5l) Yield: 63 %; M. P.:
1
-
1
-
93–195 °C; H NMR (400 MHz, DMSO-d ): d 2.31 (s,
6
H, Ar–CH ), 4.53 (s, 2H, OCH Ar), 4.65 (s, 2H, OCH ),
5-Chloro-2-hydroxy-3-{4-[(2-fluorobenzyloxy)methyl]-1H-
3
2
2
.17 (d, 2H, J = 7.6 Hz, Ar–H), 7.25 (d, 2H, J = 8.0 Hz,
1,2,3-triazol-1-yl}benzoic acid (5q) Yield: 53 %; M.P.
1
208–210 °C; H NMR (400 MHz, DMSO-d ): d 4.62 (s,
2H, OCH Ar), 4.67 (s, 2H, OCH ), 7.18–7.48 (m, 4H, Ar–
3
H), 7.73–7.74 (m, 2H, Ar–H), 8.71 (s, 1H, triazole-H);
Ar–H), 7.87 (s, 1H, Ar–H), 7.93 (s, 1H, Ar–H), 8.64 (s, 1H,
1
triazole-H); C NMR (100 MHz, DMSO-d ): d 21.2, 63.1,
6
3
6
2
2
1
7
1
1
2.0, 120.5, 125.2, 126.1, 128.5, 128.6, 128.7, 130.0,
C
30.1, 137.8, 138.5, 155.3, 170.1; IR (KBr): m 3,436, 1,636,
,471 cm ; MS (ESI): m/z 371.8 [M-H] .
NMR (100 MHz, DMSO-d ): d 63.4, 65.5, 115.6, 119.7,
120.6, 124.8, 125.4, 126.4, 128.7, 129.6, 130.8. 135.6,
6
-
1
-
123

Med Chem Res
1
44.2, 155.0, 159.5, 162.0, 170.3; IR (KBr): m 3,444, 1,639,
21.4, 46.9, 56.7, 117.8, 121.3, 126.3, 126.4, 128.3, 128.5,
128.8, 129.1, 129.8, 130.1, 131.8, 138.0, 138.5, 157.5,
-
,466, 1,420 cm ; MS (ESI): m/z 375.7 [M-H] .
1
-
1
-
1
1
69.6; IR (KBr): m 3,441, 1,637, 1,502, 1,465, 1,425 cm ;
5
1
-Chloro-2-hydroxy-3-{4-[(2-bromobenzyloxy)methyl]-
-
MS (ESI): m/z 475.2 [M-H] .
H-1,2,3-triazol-1-yl}benzoic acid (5r) Yield: 48 %;
1
M. P.: 189–191 °C; H NMR (400 MHz, DMSO-d ): d
4
5-Chloro-2-hydroxy-3-{4-[N,N-bis(2-methylbenzyl)amino]
6
.63 (s, 2H, OCH Ar), 4.75 (s, 2H, OCH ), 7.25 (t, 1H,
methyl-1H-1,2,3-triazol-1-yl}benzoic acid (5w) Yield:
1
48 %; M. P.: 132–133 °C; H NMR (400 MHz, DMSO-
2
2
J = 7.6 Hz, Ar–H), 7.40 (t, 1H, J = 8.0 Hz, Ar–H), 7.53
d, 1H, J = 7.2 Hz, Ar–H), 7.60 (d, 1H, J = 7.6 Hz, Ar–
H), 7.83 (s, 1H, Ar–H), 7.85 (s, 1H, Ar–H), 8.71 (s, 1H,
(
d ): d 2.11 (s, 6H, 2CH ), 4.02–4.15 (s, br, 6H, Ar–CH , N–
6
3
2
CH ), 7.24–7.58 (m, 8H, Ar–H), 7.81 (s, 1H, Ar–H), 7.91
2
13
(s, 1H, Ar–H), 8.95 (s, 1H, triazole-H); C NMR (DMSO-
1
3
triazole-H); C NMR (DMSO-d ): d 63.7, 71.3, 119.5,
6
1
22.7, 125.9, 126.7, 127.6, 128.2, 129.9, 132.8, 137.6,
-
d6): d 19.1, 47.6, 54.3, 119.3, 126.4, 126.7, 127.5, 128.0,
129.1, 130.0, 131.1, 131.8, 138.6, 157.6, 169.3.; IR (KBr):
-1
1
1
56.1, 169.5; IR (KBr): m 3,437, 1,638, 1,469 cm ; MS
ESI): m/z 435.7 [M-H] ; HRMS (ESI) calcd for
-
(
m 3,435, 1,640, 1,503, 1,467, 1,423 cm ; MS (ESI): m/z.
475.2 [M-H] .
-
C H BrClN O [M-H]: 435.9700, found: 435.9710.
1
7
12
3 4
5
-Chloro-2-hydroxy-3-[4-(phenylaminomethyl)-1H-1,2,3-
5-Chloro-2-hydroxy-3-{4-[N,N-bis(4-fluorobenzyl)amino]
triazol-1-yl]benzoic acid (5s) Yield: 90 %; M.P.:
methyl-1H-1,2,3-triazol-1-yl}benzoic acid (5x) Yield:
1
1
1
2
7
77–179 °C; H NMR (400 MHz, DMSO-d ): d 4.38 (s,
48 %; M. P.: 197–198 °C; H NMR (400 MHz, DMSO-
6
H, NCH ), 6.11 (s, 1H, N–H), 6.53–7.09 (m, 5H, Ar–H),
d ): d 4.13–4.15 (s, br, 6H, Ar–CH , N–CH ), 7.24–7.56
2
6
2
2
1
.75 (s, 2H, Ar–H), 8.62 (s, 1H, triazole-H); C NMR
3
(m, 8H, Ar–H), 7.79 (s, 1H, Ar–H), 7.86 (s, 1H, Ar–H),
1
3
(
DMSO-d ): d 39.0, 112.9, 116.6, 117.8, 124.2, 125.6,
8.86 (s, 1H, triazole-H); C NMR (DMSO-d ): d 46.6,
6
6
1
3
26.6, 129.3, 129.4, 146.0, 148.8, 157.1, 169.7; IR (KBr): m
,436, 1,637, 1,578, 1,507 cm ; MS (ESI): m/z 342.6
55.5, 115.9, 116.1, 118.8, 126.9, 128.2, 129.3, 130.9,
-
1
132.3, 133.2, 155.7, 161.5, 163.9, 170.2; IR (KBr): m 3,445,
1,635, 1,500, 1,465, 1,423 cm ; MS (ESI): m/z 483.1
-
-1
[
M-H] .
-
M-H] .
[
5
1
1
4
-Chloro-2-hydroxy-3-[4-(N,N-dibenzylamino)methyl-1H-
,2,3-triazol-1-yl]benzoic acid (5t) Yield: 47 %; M.P.:
1
71–172 °C; H NMR (400 MHz, DMSO-d ): d 4.21 (s,
5-Chloro-2-hydroxy-3-{4-[N,N-bis(3-fluorobenzyl)amino]
methyl-1H-1,2,3-triazol-1-yl}benzoic acid (5y) Yield:
1
52 %; M. P.: 204–205 °C; H NMR (400 MHz, DMSO-
6
H, Ar–CH ), 4.32 (s, 2H, N–CH ), 7.44–7.54 (m, 10H,
2
2
Ar–H), 7.77 (d, 1H, J = 2.8 Hz, Ar–H), 7.86 (s, 1H,
1
J = 2.8 Hz, Ar–H), 8.93 (s, 1H, triazole-H); C NMR
d ): d 4.01 (s, 4H, Ar–CH ), 4.13 (s, 2H, N–CH ),
6
2
2
3
7.20–7.65 (m, 8H, Ar–H), 7.85 (s, 1H, Ar–H), 7.94 (s, 1H,
1
3
(
DMSO-d ): d 46.8, 56.6, 118.5, 120.9, 126.4, 126.8,
Ar–H), 8.76 (s, 1H, triazole-H); C NMR (DMSO-d ): d
6
6
1
1
28.1, 129.2, 129.5, 129.9, 131.2, 138.9, 139.0, 156.8,
-
46.5, 55.6, 115.6, 116.1, 118.8, 126.9, 128.2, 129.3, 130.9,
1
69.8; IR (KBr): m 3,440, 1,638, 1,501, 1,467, 1,425 cm
;
134.7, 134.9, 155.7, 161.2, 163.3, 170.0; IR (KBr): m 3,437,
1,636, 1,503, 1,468, 1,423 cm ; MS (ESI): m/z 483.1
-
-1
MS (ESI): m/z. 447.1 [M-H] .
-Chloro-2-hydroxy-3-{4-[N,N-bis(4-methylbenzyl)amino]
methyl-1H-1,2,3-triazol-1-yl}benzoic acid (5u) Yield:
-
M-H] .
[
5
5-Chloro-2-hydroxy-3-{4-[N,N-bis(2-fluorobenzyl)amino]
1
6
6 %; M. P.: 144–146 °C; H NMR (400 MHz, DMSO-
methyl-1H-1,2,3-triazol-1-yl}benzoic acid (5z) Yield:
1
d ): d 2.32 (s, 6H, Ar–CH ), 4.18 (s, 4H, Ar–CH ), 4.29 (s,
2
66 %; M.P.: 212–213 °C; H NMR (400 MHz, DMSO-d ):
6
3
2
6
H, N–CH ), 7.26 (d, 2H, J = 7.2 Hz, Ar–H), 7.41 (d, 2H,
2
d 4.02 (s, 4H, Ar–CH ), 4.12 (s, 2H, N–CH ), 7.17–7.48
2
2
J = 7.2 Hz, Ar–H), 7.75 (s, 1H, Ar–H), 7.82 (s, 1H, Ar–
1
(m, 8H, Ar–H), 7.85 (s, 1H, Ar–H), 7.92 (s, 1H, Ar–H),
13
3
H), 8.89 (s, 1H, triazole-H); C NMR (DMSO-d ): d 21.3,
8.77 (s, 1H, triazole-H); C NMR (DMSO-d ): d 46.7,
6
6
4
1
1
6.5, 56.3, 117.8, 126.2, 126.4, 129.5, 129.7, 129.8, 130.0,
55.4, 115.7, 116.2, 118.7, 126.9, 128.3, 129.4, 130.9,
30.4, 131.3, 138.9, 139.0, 157.5, 169.5; IR (KBr): m 3,437,
,637, 1,502, 1,468, 1,424 cm ; MS (ESI): m/z 474.8
134.6, 134.8, 155.5, 161.5, 163.4, 170.1; IR (KBr): m 3,441,
1,637, 1,502, 1,465, 1,428 cm ; MS (ESI): m/z. 483.1
-
1
-1
-
?
-
[M-H] .
[
M-H] , 476.9 [M?H] .
-Chloro-2-hydroxy-3-{4-[N,N-bis(3-methylbenzyl)amino]
methyl-1H-1,2,3-triazol-1-yl}benzoic acid (5v) Yield:
5
5-Chloro-2-hydroxy-3-{4-[N,N-bis(4-nitrobenzyl)amino]
methyl-1H-1,2,3-triazol-1-yl}benzoic acid (5aa) Yield:
1
1
4
2
7
5 %; M. P.: 108–109 °C; H NMR (400 MHz, CDCl ): d
56 %; M.P.: 225–226 °C; H NMR (400 MHz, DMSO-d ):
3
6
.34 (s, 6H, 2CH ), 4.33–4.35 (s, br, 6H, Ar–CH , N–CH ),
d 3.81 (s, br, 6H, Ar–CH , N–CH ), 7.72 (d, 4H,
3
2
2
2
2
.24–7.45 (m, 8H, Ar–H), 8.13 (s, 2H, Ar–H), 9.03 (s, 1H,
1
J = 8.4 Hz, Ar–H), 7.86 (s, 2H, Ar–H), 8.20 (d, 4H,
13
J = 8.4 Hz, Ar–H), 8.65 (s, 1H, triazole-H); C NMR
3
triazole-H), 14.8 (s, 1H, OH). C NMR (DMSO-d ): d
6
1
23

Med Chem Res
(
DMSO-d ): d 47.8, 56.8, 121.4, 123.9, 126.5, 126.9,
100 lL of antidigoxigenin peroxidase (POD) was added to
each well. After incubation at 37 °C for 60 min, the plates
were washed by PBS again. The digoxigenin-labeled
products were visualized by adding tetramethylbenzidine
6
1
3
5
29.3, 130.2, 130.5, 146.7, 147.2, 154.4, 170.1; IR (KBr): m
-
1
,440, 1,638, 1,501, 1,467, 1,423 cm ; MS (ESI): m/z
-
37.1 [M-H] .
(
TMB) and a POD substrate. H SO (0.5 M, 100 lL) was
2
4
5
-Chloro-2-hydroxy-3-{4-[N,N-bis(4-bromobenzyl)amino]
added to stop the colorimetric reaction. The absorbance
methyl-1H-1,2,3-triazol-1-yl}benzoic acid (5ab) Yield:
1
6
intensity was recorded at 450 nm by a microplate reader,
0
2 %; M.P.: 135–136 °C; H NMR (400 MHz, DMSO-d ):
6
and finally total inhibitory effect (including inhibiting 3 -
d 4.02 (s, 4H, Ar–CH ), 4.13 (s, 2H, N–CH ), 7.46 (d, 4H,
J = 8.4 Hz, Ar–H), 7.62 (d, 4H, J = 8.4 Hz, Ar–H), 7.82
2
2
end process reaction and strand transfer reaction) was ob-
tained by comparing the absorbance of drug group with
that of control group (without test compound).
(
d, 1H, J = 2.8 Hz, Ar–H), 7.89 (d, 1H, J = 2.8 Hz, Ar–
1
3
H), 8.75 (s, 1H, triazole-H); C NMR (DMSO-d ): d 47.8,
6
5
1
1
6.1, 119.6, 119.7, 122.2, 126.5, 127.5, 127.8, 130.0,
Acknowledgments This work was supported by Grants from the
Beijing Natural Science Foundation (No. 7112008), and Fund to
Zeng, C. C. from Beijing City Education Committee
(KM201010005009). Zeng, C. C. thanks Ms. Chiu Marco Lam at
UCSB for the languish polishing.
32.0, 132.8, 139.4, 155.8, 170.1; IR (KBr): m 3,439, 1,636,
,503, 1,465, 1,428 cm ; MS (ESI): m/z 604.9 [M?H] .
-
1
?
5
-Chloro-2-hydroxy-3-{4-[N,N-bis(2-bromobenzyl)amino]
methyl-1H-1,2,3-triazol-1-yl}benzoic acid (5ac) Yield:
1
Conflict of interest The authors confirm that this article content has
no conflicts of interest.
5
6 %; M.P.: 175–177 °C; H NMR (400 MHz, DMSO-d ):
6
d 3.82 (s, 4H, Ar–CH ), 3.93 (s, 2H, N–CH ), 7.18–7.70
2
2
(
m, 8H, Ar–H), 7.88 (s, 1H, Ar–H), 7.97 (s, 1H, Ar–H),
1
3
8
5
1
.65 (s, 1H, triazole-H); C NMR (DMSO-d ): d 48.1,
6
4.4, 119.3, 120.4, 121.7, 127.5, 128.3, 129.5, 129.8,
References
30.2, 131.6, 133.9, 134.5, 134.6, 138.5, 157.9, 170.1; IR
-
1
Anthony NJ (2004) HIV-1 integrase: a target for new AIDS
chemotherapeutics. Curr Top Med Chem 4:979–990
(KBr): m 3,441, 1,640, 1,503, 1,470, 1,425 cm ; MS
(ESI): m/z 604.9 [M?H] .
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1
5
4 %; M.P.: 201–202 °C; H NMR (400 MHz, DMSO-d ):
6
d 3.90 (s, 4H, Ar–CH ), 3.98 (s, 2H, N–CH ), 7.28–7.71
2
2
(
m, 8H, Ar–H), 7.88 (s, 1H, Ar–H), 7.98 (s, 1H, Ar–H),
1
3
8
5
1
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6
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-
1
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respectively. The donor DNA was biotinylated at the 5 end
0
(
5
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9
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