Synthesis and Biological Evaluation of a Series of 2-((1-substituted-1H-1,2,3-triazol-4-yl)methylthio)-6-(naphthalen-1-ylmethyl)pyrimidin-4(3H)-one As Potential HIV-1 Inhibitors

Article abstract of DOI:10.1111/cbdd.12524

A series of novel S-DABO derivatives with the substituted 1,2,3-triazole moiety on the C-2 side chain were synthesized using the simple and efficient CuAAC reaction, and biologically evaluated as inhibitors of HIV-1. Among them, the most active HIV-1 inhi

Full text of DOI:10.1111/cbdd.12524

Received Date : 20-Oct-2014
Revised Date : 07-Dec-2014
Accepted Date : 09-Jan-2015
Article type : Research Article
Synthesis and biological evaluation of a series of
-((1-substituted-1H-1,2,3-triazol-4-yl)methylthio)-6-(naphthalen-1-ylm
ethyl)pyrimidin-4(3H)-one as potential HIV-1 inhibitors
2
1
#
1 #
1
1
3
Zengjun Fang , Dongwei Kang , Lingzi Zhang , Boshi Huang , Huiqing Liu , Christophe
2
2
1,*
1,*
Pannecouque , Erik De Clercq , Peng Zhan , Xinyong Liu
1
Department of Medicinal Chemistry, Key Laboratory of Chemical Biology (Ministry of
Education), School of Pharmaceutical Sciences, Shandong University, 44 West Culture Road,
2
50012 Ji’nan, Shandong, PR China;
2
Rega Institute for Medical Research, K.U.Leuven, Minderbroedersstraat 10, B-3000 Leuven,
Belgium
3
Institute of Pharmacology, School of Medicine, Shandong University, 44 West Culture Road,
2
50012 Ji’nan, Shandong, PR China
*
Corresponding authors. Tel: +86 531 88380270; fax: +86 531 88382731; E-mail address:
zhanpeng1982@163.com (Zhan P.); xinyongllab@163.com (Liu X.Y.)
This article has been accepted for publication and undergone full peer review but has
not been through the copyediting, typesetting, pagination and proofreading process
which may lead to differences between this version and the Version of Record. Please
cite this article as an 'Accepted Article', doi: 10.1111/cbdd.12524
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#Contributed equally to this work
Abstract: A series of novel S-DABO derivatives with the substituted 1,2,3-triazole
moiety on the C-2 side chain were synthesized by using the simple and efficient
CuAAC reaction, and biologically evaluated as inhibitors of HIV-1. Among them, the
most active HIV-1 inhibitors was compound
4
1
-((4-((4-(2,6-dichlorobenzyl)-5-methyl-6-oxo-1,6-dihydropyrimidin-2-ylthio)methyl)-
H-1,2,3-triazol-1-yl)methyl)benzenesulfonamide (B5b7), which exhibited similar
HIV-1 inhibitory potency (EC = 3.22 μM) compared with 3TC (EC = 2.24 μM). None
5
0
50
of these compounds demonstrated inhibition against HIV-2 replication. The
preliminary structure-activity relationship (SAR) of these new derivatives was
discussed briefly.
Keywords: HIV, NNRTIs, Pyrimidine, Click chemistry, Antiviral activity, Drug design,
SAR
1
. Introduction
Human immunodeficiency virus type-1 (HIV-1) non-nucleoside reverse transcriptase
inhibitors (NNRTIs) were the major components of highly active antiretroviral
therapy (HAART), a major problem is that the long-term efficacy of NNRTIs is limited
by the rapid emergence of drug-resistant HIV-1 strains. Thus, in recent years, the
discovery of novel NNRTIs chemical entities with improved drug resistance profiles
was considered to be a very active research field [1-3].
Dihydro-alkoxy-benzyl-oxopyrimidines (DABOs) were one of the most
representative classes of HIV-1 NNRTIs developed in the past two decades. Up to
now, three generations of DABO analogues have been investigated on the base of
the C-2 substitutent features, i.e.: dihydro-alkyloxy-benzyl-oxopyrimidines
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(
O-DABOs), dihydro-alkylthio- benzyl-oxopyrimidines (S-DABOs) and
dihydro-alkylamino-difluorobenzyloxopyrimidines (N-DABOs) (Figure 1) [4].
Among DABO derivatives, S-DABOs were the most prominent anti-HIV agents,
with the C-2 alkylthio chain being extremely important for exhibition of anti-HIV-1
activity. The NHC=O fragment at N3/C4 position of the pyrimidine is a critical factor
for the binding because the key hydrogen bonds were formed between N3-H and the
carbonyl group of Lys101 [5]. The optimal moieties at positions 5 of the pyrimidine
nucleus were dependent on the nature of the C-2 side chain. Both
arylcarbonylmethyl and benzyl substituent of S-DABOs at the C-2 site of the
pyrimidine core exert their potent anti-HIV-1 effects by the interaction with the side
chain of Pro236 [6]. Various chemical modifications of C-6 position had been carried
out in the lab of Artico [7]. When the benzyl group at C-6 of the pyrimidine ring was
replaced by 1/2-naphthylmethyl or halogenated benzyl moieties, a set of compounds
demonstrated favorable potencies against wild-type (WT) HIV-1 in the
low-micromolar range. The introduction of the hydrophobic C-6 side chain of
S-DABOs might be more favorable to improve a putative π-stacking interaction
between the aryl ring of the ligand and Tyr188 or Tyr181 of RT [8].
Figure 1. The structures of DABOs families.
Figure 2. The newly designed S-DABOs with the substituted triazole moiety on the C2 side
chain.
Recent work in our lab on S-DABOs reported that assembly of a
phenylaminocarbonylmethylthio moiety at the C2 site of the pyrimidinecore has led
to a set of novel lead compounds with promising antiviral potencies against HIV-1
(
(
Figure 2) [9a,b]. Among them, MY-4b3 was identified as the most potent molecule
EC50 = 0.18 μM, CC50 >243.56μM, SI >1326). Moreover, a novel C-2 thioethanone
S-DABO derivative JZ-6c1 (EC50=0.24 μM) was reported to demonstrate the improved
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or comparable HIV-1 potency compared with NVP (EC50=0.21 μM) and DLV
EC =0.32 μM) [10]. As the C-2 side chain of S-DABOs pointed toward the solvent
(
5
0
exposed region and accommodated substantial modifications of the inhibitor, we
postulated that the introduction of the thioacetanilide and thioethanone group to
the C-2 side chain might be more favorable to improve specific protein-inhibitor
interactions, and further exploration of the plasticity of this part of the pocket may
lead to the discovery of new S-DABOs with improved pharmacokinetic properties
without a significant loss of binding affinity. These interesting clues prompted us to
further explore the structure-activity relationships (SARs) of the C-2 side chain in
S-DABOs.
In recent years, the 1,2,3-triazole structural motif, the product obtained from the
Cu(I)-catalyzed, alkyne-azide 1,3-dipolar cycloaddition (CuAAC) reaction, has
emerged as attractive and desirable scaffolds in the development of potential drug
molecules in medicinal chemistry in this regard considering the fact that the triazole
is a safe bioequivalent surrogate for the amide bond, which demonstrates high
dipole and enhanced hydrogen bonding interactions compared to the amide.
Because of their chemical features, these triazoles were proposed to be aggressive
pharmacophores that participate in drug-receptor interactions while maintaining an
excellent chemical and metabolic profile [11].
Based on the existing SARs of C2-functionalized S-DABOs [12], we proposed that the
structural modification of S-DABOs by incorporating the substituted triazole moiety
to the corresponding site of C-2 side chain could enhance the interactions between
RT and the inhibitors. In order to examine our proposal and generate more potent
NNRTIs, in the present work, a series of novel S-DABO derivatives with the
substituted 1,2,3-triazole moiety on the C-2 side chain were designed, synthesized by
using the simple and efficient CuAAC reaction, and evaluated for their anti-HIV
activities (Figure 2).
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2
. Results and discussions
2
.1. Chemistry
The synthetic route of the newly designed compounds is described in Scheme 1. The
key intermediate β-ketoesters (A2a, A2b, B2a, B2b) were prepared with a simple
method previously used by us [10] through the reaction of 2-(naphthalen-1-yl)acetic
acid (A1) or 2-(2,6-dichlorophenyl)acetic acid (B1) with N,N′-carbonyldiimidazole
(
CDI) followed by treatment with potassium 3-ethoxy-3-oxopropanoate (a) or
potassium 3-ethoxy-2-methyl-3-oxopropanoate (b) in the presence of anhydrous
MgCl and Et N. Next, the cyclization reaction of β-ketoesters (A2a, A2b, B2a, B2b)
2
3
with thiourea in the presence of EtONa (in refluxing EtOH) afforded the substituted
uracils (A3a, A3b, B3a, B3b). Treatment of these uracils with 3-bromoprop-1-yne in
the presence of K CO in anhydrous DMF gave the key alkyne intermadeties (A4a,
2
3
A4b, B4a, B4b). Then, the synthesis of these triazole linked S-DABOs (A5a1-A5a7,
A5b1-A5b7, B5a1-B5a7 and B5b1-B5b7) was accomplished using the copper
catalyzed azido-alkyne cycloaddition (CuAAC) reaction in the mild conditions. The
substituted benzyl azides were easily synthesized by the reaction of substituted
benzyl halide with sodium azide at room temperature. Finally, the iodination of
compounds A5a1-A5a7 and B5a1-B5a7 occurred subsequently to prepare
compounds A6a1-A6a7 and B6a1-B6a7, respectively, using PbO and I in HOAc.
2
2
Both analytical and spectral data of all the title derivatives are in full agreement with
the proposed structures.
Scheme 1. Reagents and Conditions: (i) (a) MgCl
2
, Et
3
N, CH CN, rt, 2h; (b) CDI, rt, overnight
3
then reflux, 2 h; (ii) thiourea, EtONa, EtOH, reflux, 6h; (iii) HC≡CCH
Br, K
CO , DMF; (iv)
2
2
3
1
(
substituted)Ar -CH
2
N
3
, VcNa, CuSO
4
.5H
2
O, H
2
O, tertiary butanol, 65°C; (v) HOAc, PbO
2
2
, I .
2
.2. Anti-HIV activity evaluation
The newly synthesized S-DABO derivatives were evaluated for their cytotoxicity
and anti-HIV activities in MT-4 cell cultures infected by HIV-1 IIIB strain (WT), HIV-1
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mutant strain RES056 (K103N/Y181C double mutanted RT) and HIV-2 strain ROD,
respectively. The FDA-approved drugs nevirapine (NVP), lamivudine (3TC),
azidothymidine (AZT), Didanosine (DDI), delavirdine (DLV), efavirenz (EFV) and
etravirine (ETV, TMC125) were chosen as reference drugs. Comparisons of the
antiviral inhibitory concentration (EC ), cytotoxic concentration (CC ), and SI
5
0
50
(
selectivity, given by the CC50/EC50 ratio) values for diverse compounds were
depicted in Table 1 and Table 2.
Table 1.
5
4
-Hydrogen/methyl/iodo-6-(naphthalen-1-ylmethyl)-2-((1-(arylmethyl)-1H-1,2,3-triazol-
-yl)methylthio)pyrimidin-4(3H)-one.
Table 2. 5-Hydrogen/methyl/iodo-6-(2,6-dichlorobenzyl)-2-((1-(arylmethyl)-1H-1,2,3-triazol-
-yl)methylthio)pyrimidin-4(3H)-one.
4
The results revealed that six target compounds (B5b1, B5B3-B5b7) had remarkable
activities against WT HIV-1, with EC50 values in the low micromolecular range, which
was better than that of DDI (EC50 = 23.20 μM). Even so, they were less active in
comparison with the previously reported C-2 substituted S-DABO derivatives and
other reference drugs [9,10]. The most active compound
4
1
-((4-((4-(2,6-dichlorobenzyl)-5-methyl-6-oxo-1,6-dihydropyrimidin-2-ylthio)methyl)-
H-1,2,3-triazol-1-yl)methyl)benzenesulfonamide (B5b7) showed activity with EC50
value of 3.22 μM comparable to that of 3TC (EC = 2.24 μM). In addition, these
5
0
S-DABOs analogues did not proved active HIV-2 and HIV-1 mutant strain RES056.
Taking together, these data we can assume that the novel S-DABOs most probably
act as genuine NNRTIs.
In terms of SARs, it became immediately apparent that substitution at C-6 of
the pyrimidine ring were detrimental, since the 2,6-dichlorobenzyl subseries was
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generally more active than the corresponding naphthalen-1-ylmethyl subseries. Only
compound A5b7 in naphthalen-1-ylmethyl series demonstrated weak HIV inhibitory
effect.
As with the SAR of previous S-DABOs, the presence of suitable substituents at
the C-5 position of uracil ring closely correlated with the antiviral activity. Optimal
anti-HIV-1 activity was obtained with compounds bearing a methyl moiety at the C-5
site (B5b1-B5b7). There was still one compound (B5a1) in C-5 unsubstituted
subseries (B5a1-B5a7) with EC50 value in double-digit mrcromolar level. Whereas,
when a iodine atom was introduced to this position, the bioactivity was completely
impaired.
Moreover, this study also suggested that the nature of the substituent on the
4
-position of the phenyl ring (terminal of C-2 side chain) significantly influenced on
the antiviral activity of these novel S-DABOs. Notably, among compounds
B5b1-B5b7, SO NH (B5b7) appeared to be the most favorable group, closely
2
2
followed in the sequence 4-COMe > 4-OMe > 4-CN > 4-CONH > 4-H > 4-F. It is
2
interesting to note that, in many types of NNRTIs, the optimum activity was often
found with substituents containing hydrophilic SO
2
NH , indicating its nature can
2
better accommodate the chemical environment in this region of RT and provide
potential interactions with amino acids. This conclusion was consistent with those
previously observed results in the arylazolyl(azinyl)thioacetanilides series [13,14].
3
. Conclusion
As part of a project devoted to structural optimization of the S-DABOs NNRTIs
and to the further elucidation of the SARs around this scaffold, a new series of
S-DABO analogues, characterized by the same central scaffold and a variously
functionalized 1,2,3-triazole linked side chain at C-2 position of the pyrimidone ring,
were prepared and assessed for their anti-HIV activity and cytotoxicity in vitro.
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Generally, some of the synthesized compounds displayed remarkable anti-HIV-1
WT) activity in low micromolar level. The best compound of this series was
(
compound B5b7, which exhibited potent activity with EC50 value of 3.22 μM
comparable to that of 3TC (EC50 = 2.24 μM). Preliminary SAR studies provided some
insights for discovery of more potent NNRTIs.
4
4
. Experimental Section
.1 Chemistry
All melting points were determined on a micro melting point apparatus and are
1
13
uncorrected. H-NMR and C-NMR spectra were obtained on a Bruker
Avance-400NMR-spectrometer in the indicated solvents. Chemical shifts are
expressed in δ units and TMS as internal reference. Mass spectra were taken on a LC
Autos ampler Device: Standard G1313A instrument. TLC was performed on Silica Gel
GF254 for TLC (Merck) and spots were visualized by iodine vapours or by irradiation
with UV light (λ= 254 nm). Flash column chromatography was performed on column
packed with Silica Gel60 (230-400 mesh). Solvents were reagent grade, when
necessary, were purified and dried by standard methods. Concentration of the
reaction solutions involved the use of rotary evaporator at reduced pressure.
4
.1.1 General procedure for the preparation of substituted benzyl azides
Different substituted benzyl halide (1 mmol) were dissolved in anhydrous DMF (10
mL), followed by addition of sodium azide (1.2 mmol). The reaction mixture was
stirred at room temperature overnight. Then 50 mL water was poured into the
mixture, the mixture was extracted with ethyl acetate (3×10 mL). The combined
organic layer was dried over anhydrous Na SO , filtered and concentrated to give the
2
4
corresponding substituted benzyl azides as crude products.
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4
5
.1.2 General procedure for the preparation of
-hydrogen/methyl/iodo-6-(naphthalen-1-ylmethyl)-2-((1-(arylmethyl)-1H-1,2,3-tri
azol-4-yl)methylthio)pyrimidin-4(3H)-ones.
General procedure for the preparation of ethyl 4-(naphthalen-1-yl)-3-oxobutanoate
(
A2a) or ethyl 2-methyl-4-(naphthalen-1-yl)-3-oxobutanoate (A2b)
A mixture solution of starting material potassium 3-ethoxy-3-oxopropanoate (a) or
potassium 3-ethoxy-2-methyl-3-oxopropanoate (b) (75.2 mmol), MgCl (9.0 g, 94.5
mmol), Et N (12.1 g, 120 mmol) in 150 mL anhydrous CH CN were stirred at room
2
3
3
temperature for 2 hours. Then this solution were added to another mixture solution
of 2-(naphthalen-1-yl)acetic acid (7.0 g, 37.6 mmol) and CDI (7.3 g, 45 mmol) in 100
mL anhydrous CH CN. The reaction mixture was stirred overnight at room
3
temperature and then heated to reflux for 2h. After the mixture was cooled, a
solution of 3N HCl (100 mL) was added slowly while the temperature was kept below
2
0°C, and the resulting clear mixtures were stirred for another 20 minutes. The
organic layer was separated and concentrated, and the residue was treated with
EtOAc (80 mL). The aqueous layer was extracted with EtOAc (3×100 mL), and the
combined organic layers were washed with saturated NaHCO
3×100 mL), dried over anhydrous Na SO , filtered and concentrated to give the
crude product ethyl 4-(naphthalen-1-yl)-3-oxobutanoate (A2a) or ethyl
-methyl-4-(naphthalen-1-yl)-3-oxobutanoate (A2b) as yellow oil, respectively, which
3
(3×100 mL) and brine
(
2
4
2
were directly used in the next step without further purification.
General procedure for the preparation of
6
5
-(naphthalen-1-ylmethyl)-2-thioxo-2,3-dihydropyrimidin-4(1H)-one (A3a) or
-methyl-6-(naphthalen-1-ylmethyl)-2-thioxo-2,3-dihydropyrimidin-4(1H)-one (A3b)
Sodium metal (0.92 g, 40mmol) was dissolved in absolute ethanol (20 mL), then
thiourea (2.28 g, 30mmol) and the intermediate A2a or A2b (10 mmol) was added
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successively. The reaction mixture was heated in reflux for 6 hours, then was
evaporated to dryness under reduced pressure. The residue was re-dissolved in H O
2
(
50 mL). The solution was acidified with 3M HCl solution to pH 4. The resulting
precipitate was filtered, washed with H O and Et O successively, and dried to give
2
2
the crude product A3a or A3b as white solid, which was directly used in the following
step without further purification.
6
-(naphthalen-1-ylmethyl)-2-thioxo-2,3-dihydropyrimidin-4(1H)-one(A3a)
1
H NMR (400 MHz, DMSO-d6, ppm) δ: 12.60 (1H, s, NH), 12.42 (1H, s, NH),
7
5
C
.95-7.98 (2H, m, aromatic), 7.89 (1H, d, J = 8.00 Hz), 7.48-7.58 (4H, m, aromatic),
.13 (1H, s, CO-CH), 4.23(2H, s, Ph-CH
2
). ESI-MS: m/z 269.4 (M+1), 291.3 (M+Na).
15
H
N
12 2
OS (268.07).
5
-methyl-6-(naphthalen-1-ylmethyl)-2-thioxo-2,3-dihydropyrimidin-4(1H)-one
(A3b)
1
H NMR (400 MHz, DMSO-d6, ppm) δ: 12.62 (1H, s, NH), 12.43 (1H, s, NH), 7.95-7.99
2H, m, aromatic), 7.88 (1H, d, J = 8.00 Hz), 7.47-7.58 (4H, m, aromatic), 4.28(2H, s,
Ph-CH ), 1.39(3H, s, CH ). ESI-MS: m/z 283.3 (M+1), 305.4 (M+Na). C16 OS
282.08).
(
2
3
H
N
14 2
(
General procedure for the preparation of
6
5
-(naphthalen-1-ylmethyl)-2-(prop-2-ynylthio)pyrimidin-4(3H)-one (A4a) or
-methyl-6-(naphthalen-1-ylmethyl)-2-(prop-2-ynylthio)pyrimidin-4(3H)-one (A4b)
A mixture solution of intermediate A3a or A3b (7mmol), K
2
CO (7mmol, 0.96 g) in 30
3
mL anhydrous DMF was stirred at room temperature for 30 min. Then propargyl
bromide (6.3 mmol, 0.74 g) in 5 mL anhydrous DMF were dropwise added into the
solution in 4 hours. TLC analysis indicated that the reaction was finished in 15h. Then
1
00 mL ice water was poured into the mixture. The formed solid was filtrated,
washed with water. Then the crude product was recrystallized in MeOH to get the
desired product A4a or A4b as yellow solid.
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6
-(naphthalen-1-ylmethyl)-2-(prop-2-ynylthio)pyrimidin-4(3H)-one(A4a)
1
H NMR (400 MHz, DMSO-d6, ppm) δ: 12.74 (1H, s, NH), 7.95-7.98 (2H, m, aromatic),
.91 (1H, d, J = 8.00 Hz), 7.48-7.59 (4H, m, aromatic), 5.13 (1H, s, CO-CH), 4.23(2H, s,
Ph-CH ), 3.96(2H, s, S-CH ), 3.08(1H, s, C≡CH). ESI-MS: m/z 307.3 (M+1), 329.4
M+Na). C18 OS (306.08).
7
2
2
(
H
N
14 2
5
-methyl-6-(naphthalen-1-ylmethyl)-2-(prop-2-ynylthio)pyrimidin-4(3H)-one (A4b)
1
H NMR (400 MHz, DMSO-d6, ppm) δ: 12.74 (1H, s, NH), 7.96-7.98 (2H, m, aromatic),
.89 (1H, d, J = 8.00 Hz), 7.48-7.57 (4H, m, aromatic), 4.20 (2H, s, Ph-CH ), 3.91(2H,
s, S-CH ). ESI-MS: m/z 321.3 (M+1), 343.5
), 3.07(1H, s, C≡CH), 1.39 (3H, s, CH
M+Na). C19 OS (320.10).
7
2
2
3
(
H
N
16 2
General procedure for the preparation of
-(naphthalen-1-ylmethyl)-2-((1-(arylmethyl)-1H-1,2,3-triazol-4-yl)methylthio)pyri
midin-4(3H)-one (A5a1-A5a7) or
-methyl-6-(naphthalen-1-ylmethyl)-2-((1-(arylmethyl)-1H-1,2,3-triazol-4-yl)methyl
6
5
thio)pyrimidin-4(3H)-one (A5b1-A5b7)
To a mixture of intermediate A4a or A4b (0.65 mmol) and substituted benzyl azide
(
0.65 mmol) in water and tertiary butanol (v/v= 1:1, 15 mL), the water solution (1N)
of sodiumascorbate (0.04 g, 0.195 mmol) was added, followed by the addition of
copper (II) sulfate pentahydrate (0.065 mmol, 0.017g) in water (7.5 % solution). The
heterogeneous mixture was stirred vigorously at 65°C for 8 hours. The formed solid
was filtrated and washed with water. Then the crude product was recrystallized in
MeOH to give the products
6
-(naphthalen-1-ylmethyl)-2-((1-(arylmethyl)-1H-1,2,3-triazol-4-yl)methylthio)pyrimi
din-4(3H)-one (A5a1-A5a7) or
-methyl-6-(naphthalen-1-ylmethyl)-2-((1-(arylmethyl)-1H-1,2,3-triazol-4-yl)methylth
5
io)pyrimidin-4(3H)-one (A5b1-A5b7) as white or light yellow solids.
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General procedure for the preparation of
-iodo-6-(naphthalen-1-ylmethyl)-2-((1-(arylmethyl)-1H-1,2,3-triazol-4-yl)methylthi
5
o)pyrimidin-4(3H)-one (A5c1-A5c7)
To a solution of A5a1-A5a7 (0.5 mmol) in HOAc (5 mL) was added the PbO (0.30
2
mmol) and I (0.30 mmol), and the mixture were stirred at room temperature. After
2
completion of the reaction according to TLC analysis, the mixture was poured into
H
2
O (50 mL). The resulting solid was then filtered, washed with water, and purified
by silica gel column chromatography to afford the final products A5c1-A5c7.
2
-(((1-(4-fluorobenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(naphthalen-1-ylmethyl)
pyrimidin-4(3H)-one (A5a1)
1
Recrystallized from methanol as a light yellow crystal, yield 72.1%, mp:195-197°C. H
NMR (400 MHz, DMSO-d6, ppm) δ: 12.54 (1H, s, NH), 8.11 (1H, dd, J = 5.60, 3.24 Hz),
7
.82 (1H, d, J = 7.60 Hz), 7.66 (1H, s, triazole-H), 7.44-7.51 (4H, m, aromatic), 7.30
2H, dd, J = 8.52, 5.72 Hz), 7.19 (2H , t, J = 8.88 Hz), 5.92 (1H, s, CO-CH), 5.47 (2H , s,
Ph-CH -triazole), 4.31 (2H, s, Ph-CH ), 4.30 ( 2H, s, S-CH ). ESI-MS:m/z 458.4 (M+1),
80.3 (M+Na). C25 OS (457.52).
(
2
2
2
4
H
20FN
5
2
-(((1-benzyl-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(naphthalen-1-ylmethyl)pyrimidin
-
4(3H)-one (A5a2)
1
Recrystallized from methanol as a white crystal, yield 63.7%, mp:190-192°C. H NMR
(
(
(
400 MHz, DMSO-d6, ppm) δ: 12.54 (1H, s, NH), 8.11 (1H, dd, J = 5.92, 3.16 Hz), 7.91
1H, dd, J = 6.08, 3.24 Hz), 7.81 (1H, d, J = 7.68 Hz), 7.64 (1H, s, triazole-H), 7.43-7.51
4H, m, aromatic), 7.30-7.37 (3H, m, aromatic), 7.21 (2H, d, J = 6.84 Hz), 5.92 (1H, s,
CO-CH), 5.47 (2H, s, Ph-CH
2
-triazole), 4.31 (2H, s, Ph-CH
2
), 4.29 (2H, s, S-CH ).
2
ESI-MS:m/z 440.4 (M+1), 462.4(M+Na). C25
H
N
21 5
OS (439.53).
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2
-(((1-(4-methoxybenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(naphthalen-1-ylmeth
yl)pyrimidin-4(3H)-one (A5a3)
1
Recrystallized from methanol as a white crystal, yield 70.2%, mp:193-195°C. H NMR
(
(
(
400 MHz, DMSO-d6, ppm) δ:12.56 (1H, s, NH), 8.11 (1H, dd, J = 5.68, 3.00 Hz), 7.92
1H, dd, J = 6.08, 3.24 Hz), 7.83 (1H, d, J = 7.84 Hz), 7.60 (1H, s, triazole-H), 7.44-7.51
4H, m, aromatic), 7.20 (2H, d, J = 8.40 Hz), 6.89 (2H, d, J = 8.48 Hz),5.93 (1H, s,
CO-CH), 5.39 (2H, s, Ph-CH
2
-triazole), 4.30 (2H, s, Ph-CH
). C NMR (100 MHz, DMSO-d6, ppm) δ: 159.6, 134.4, 133.9, 132.1,
29.9, 128.9, 128.5, 128.3, 127.8, 126.6, 126.2, 126.1, 124.9, 123.5, 114.5, 55.6, 52.7,
4.8. ESI-MS: m/z 470.4 (M+1), 492.3 (M+Na). C26 S (469.56).
2
), 4.29 (2H, s, S-CH
2
), 3.73
1
3
(
3H, s, OCH
3
1
2
H
23
N
O
5 2
4
-((4-(((4-(naphthalen-1-ylmethyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)-1H
-
1,2,3-triazol-1-yl)methyl)benzamide (A5a4)
1
Recrystallized from methanol as a white crystal, yield 75.4%, mp:250-252°C. H NMR
(
(
400 MHz, DMSO-d6, ppm) δ: 12.54 (1H, s, NH), 8.11 (1H , d, J = 4.52 Hz), 7.80-7.98
5H, m, aromatic), 7.67 (1H, s, triazole-H), 7.40-7.50 (5H, m, aromatic), 7.27 (2H, d, J
=
4
1
5
7.80 Hz), 5.94 (1H, s, CO-CH), 5.55 (2H, s, Ph-CH
2
-triazole), 4.33 (2H, s, Ph-CH
2
),
1
3
.30 (2H, s, S-CH
2
). C NMR (100 MHz, DMSO-d6, ppm) δ: 167.9, 139.4, 134.6, 134.4,
33.9, 132.1, 128.9, 128.4, 128.3, 127.9, 127.8, 126.6, 126.2, 126.1, 124.9, 124.0,
2.8, 24.9. ESI-MS: m/z 483.3 (M+1), 505.3(M+Na). C H N O S (482.56).
2
6 22 6 2
2
-((4-(((4-(naphthalen-1-ylmethyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)-1H
-
1,2,3-triazol-1-yl)methyl)benzonitrile (A5a5)
1
Recrystallized from methanol as a white crystal, yield 67.2%, mp: 170-173°C. H NMR
(
(
(
400 MHz, DMSO-d6, ppm) δ: 12.59 (1H, s, NH), 8.11 (1H , dd, J = 8.92, 3.16 Hz), 7.92
2H , dd, J = 12.40, 7.16 Hz), 7.77-7.82 (2H, m, aromatic), 7.70(1H, t, J = 7.48 Hz), 7.56
1H, t, J = 7.52 Hz), 7.44-7.50 (4H, m, aromatic), 7.22 (1H, d, J = 7.76 Hz), 5.91 (1H, s,
CO-CH), 5.71 (2H, s, Ph-CH
2
-triazole), 4.33 (2H, s, Ph-CH
OS (464.54).
2
), 4.31 (2H, s, S-CH ). ESI-MS:
2
m/z 465.3 (M+1), 487.3 (M+Na). C26
H
20 6
N
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2
-(((1-(2-methylbenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(naphthalen-1-ylmethyl
)pyrimidin-4(3H)-one (A5a6)
1
Recrystallized from methanol as a white crystal, yield 62.1%, mp:101-104°C. H NMR
(
(
(
(
400 MHz, DMSO-d6, ppm) δ:12.54 (1H, s, NH), 8.10 (1H, dd, J = 9.56, 3.44 Hz), 7.88
1H, d, J = 3.32 Hz), 7.79 (1H, dd, J = 8.60, 1.20 Hz), 7.58 (1H, s, triazole-H), 7.42-7.52
4H, m, aromatic), 7.24 (1H, td, J = 7.44, 0.72 Hz), 7.16 (1H, td, J = 7.48, 2.32 Hz), 6.93
1H, d, J = 7.56 Hz), 5.92 (1H, s, CO-CH), 5.53 (2H, s, Ph-CH
), 4.28 (2H, s, S-CH ), 2.27 (3H, s, Ph-CH ). ESI-MS: m/z 454.3 (M+1) , 476.3
M+Na). C H N OS (453.56).
2
-triazole), 4.31 (2H, s,
Ph-CH
2
2
3
(
2
6 23 5
4
-((4-(((4-(naphthalen-1-ylmethyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)-1H
-
1,2,3-triazol-1-yl)methyl)benzenesulfonamide (A5a7)
1
Recrystallized from methanol as a white crystal, yield 79.3%, mp:140-142°C. H NMR
(
(
400 MHz, DMSO-d6, ppm) δ: 12.49 (1H, s, NH), 8.13 (1H, dd, J = 9.48, 3.64 Hz), 7.91
1H, dd, J = 5.52, 2.04 Hz), 7.80-7.83 (4H, m, aromatic), 7.67 (1H, s, triazole-H),
7
.30-7.52 (5H, m, aromatic), 5.93 (1H, s, CO-CH), 5.56 (2H, s, Ph-CH
2H, s, Ph-CH ), 4.30 (2H, s, S-CH ). ESI-MS: m/z 519.3 (M+1), 541.3 (M+Na).
(518.61).
2
-triazole), 4.32
(
2
2
C
25
H
22
N
6
O
3 2
S
2
-(((1-(4-fluorobenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-5-methyl-6-(naphthalen-1-
ylmethyl)pyrimidin-4(3H)-one (A5b1)
1
Recrystallized from methanol as a light yellow crystal, yield 75.6%, mp: 207-210°C. H
NMR (400 MHz, DMSO-d6, ppm) δ: 12.52 (1H, s, NH), 8.13 (1H, dd, J = 5.96, 2.48 Hz),
7
.88 (1H, dd, J = 6.68, 3.96 Hz), 7.75 (1H, d, J = 8.16 Hz), 7.49 (2H, td, J = 6.04, 2.32
Hz), 7.38 (1H, t, J = 7.24 Hz), 7.15-7.29 (6H, m, aromatic), 5.38 (2H, s,
Ph-CH -triazole), 4.36 (2H, s, Ph-CH ), 4.02 (2H, s, S-CH ), 2.03 (3H, s,
2
2
2
pyrimidone-CH
3
). ESI-MS: m/z 472.4 (M+1), 494.5 (M+Na). C26
H
22FN OS (471.55).
5
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2
-(((1-benzyl-1H-1,2,3-triazol-4-yl)methyl)thio)-5-methyl-6-(naphthalen-1-ylmethyl)
pyrimidin-4(3H)-one (A5b2)
1
Recrystallized from methanol as a white crystal, yield 61.0%, mp:175-177°C. H NMR
(
(
400 MHz, DMSO-d6, ppm) δ: 12.61 (1H, s, NH), 8.13 (1H, dd, J = 9.12, 1.80 Hz), 7.87
1H, dd, J = 7.00, 2.92 Hz), 7.73 (1H, d, J = 8.16 Hz), 7.47-7.51 (2H, m, aromatic),
7
.25-7.39 (6H, m, aromatic), 7.16 (2H, dd, J = 7.64, 2.20 Hz), 5.38 (2H, s,
Ph-CH -triazole), 4.36 (2H, s, Ph-CH ), 4.04 (2H, s, S-CH ), 2.04 (3H, s,
pyrimidone-CH ). ESI-MS: m/z 454.4 (M+1), 476.4 (M+Na). C26
2
2
2
3
H
23
N
5
OS (453.56).
2
-(((1-(4-methoxybenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-5-methyl-6-(naphthalen
-
1-ylmethyl)pyrimidin-4(3H)-one (A5b3)
1
Recrystallized from methanol as a white crystal, yield 72.4%, mp:164-166°C. H NMR
(
(
400 MHz, DMSO-d6, ppm) δ: 12.56 (1H, s, NH), 8.15 (1H, dd, J = 9.16, 2.28 Hz), 7.89
1H, dd, J = 6.68, 3.92 Hz), 7.76 (1H, d, J = 8.16 Hz), 7.50 (2H, td, J = 6.04, 2.32 Hz),
7
.39 (1H, t, J = 7.32 Hz), 7.26 (1H, d, J =5.12 Hz), 7.12 (1H, d, J = 8.60 Hz), 6.87 (2H, d,
J = 8.64 Hz), 5.30 (2H, s, Ph-CH -triazole), 4.34 (2H, s, Ph-CH ), 4.04 (2H, s, S-CH ),
2
2
2
1
3
3
.73 (3H, s, O-CH
3
), 2.02 (3H, s, pyrimidone-CH ). C NMR (100 MHz, DMSO-d6,
3
ppm) δ: 159.5, 144.0, 135.0, 133.8, 132.4, 129.8, 128.8, 128.2, 127.3, 127.1, 126.4,
26.2, 126.0, 124.8, 123.1, 114.5, 55.6, 52.7, 37.7, 24.5, 11.0. ESI-MS: : m/z 484.5
M+1), 506.5(M+Na). C H N O S (483.58).
1
(
2
7 25 5 2
4
-((4-(((5-methyl-4-(naphthalen-1-ylmethyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)
methyl)-1H-1,2,3-triazol-1-yl)methyl)benzamide (A5b4)
1
Recrystallized from methanol as a white crystal, yield 63.2%, mp: 238-240°C. H NMR
400 MHz, DMSO-d6, ppm) δ: 12.54 (1H, s, NH), 8.12 (1H, d, J = 7.56 Hz), 7.95 (1H, s),
(
7
7
.83-7.88 (3H, m, aromatic), 7.73 (1H, d, J = 8.12 Hz), 7.48 (2H, td, J = 5.04, 1.64 Hz),
.37 (2H, t, J =7.36 Hz), 7.27-7.30 (2H, m, aromatic), 7.20 (2H, d, J = 8.16 Hz), 5.45
(
2H, s, Ph-CH
2
-triazole), 4.35 (2H, s, Ph-CH
2
), 4.05 (2H, s, S-CH
2
), 2.02 (3H, s,
13
pyrimidone-CH
3
). C NMR (100 MHz, DMSO-d6, ppm) δ:167.8, 144.1, 139.4, 134.9,
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1
1
34.5, 133.7, 132.4, 128.8, 128.3, 127.9, 127.3, 127.2, 126.4, 126.1, 125.9, 124.8,
23.6, 52.7, 37.6, 24.4, 11.0. ESI-MS: m/z 497.5 (M+1), 519.4(M+Na). C H N O S
2
7 24 6 2
(
496.58).
2
-((4-(((5-methyl-4-(naphthalen-1-ylmethyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)
methyl)-1H-1,2,3-triazol-1-yl)methyl)benzonitrile (A5b5)
1
Recrystallized from methanol as a yellow crystal, yield 71.0%, mp: 130-132°C. H
NMR (400 MHz, DMSO-d6, ppm) δ:12.62 (1H, s, NH), 8.12 (1H, dd, J = 8.92, 3.16 Hz),
7
7
.92 (2H, dd, J = 10.60, 7.16 Hz), 7.62-7.82 (2H, m, aromatic), 7.70 (1H, t, J = 7.62 Hz),
.52 (1H, t, J = 7.52 Hz), 7.42-7.53 (2H, m, aromatic), 7.20 (1H, d, J = 7.76 Hz), 7.14
(
1H, d, J = 7.76 Hz), 5.61 (2H, s, Ph-CH
2
-triazole), 4.36 (2H, s, Ph-CH
2
), 4.06 (2H, s,
S-CH
2
), 2.04 (3H, s, pyrimidone-CH
3
). ESI-MS: m/z 479.4 (M+1), 501.4 (M+Na).
C H N OS (478.57).
2
7 22 6
5
-methyl-2-(((1-(2-methylbenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(naphthalen-1
-
ylmethyl)pyrimidin-4(3H)-one (A5b6)
1
Recrystallized from methanol as a white crystal, yield 72.4%, mp:145-147°C. H NMR
400 MHz, DMSO-d6, ppm) δ: 12.54 (1H, s, NH), 8.10 (1H, d, J = 7.92 Hz), 7.84 (1H,
dd, J = 7.28, 1.72 Hz), 7.71 (1H, d, J = 8.16 Hz), 7.49 (2H, td, J = 7.68, 1.40 Hz), 7.35
(
(
(
1H, t, J = 7.28 Hz), 7.21-7.27 (4H, m, aromatic), 7.12 (1H, td, J = 7.48, 1.60 Hz), 6.84
1H, d, J = 7.52 Hz), 5.40 (2H, s, Ph-CH -triazole), 4.33 (2H, s, Ph-CH ), 4.04 (2H, s,
), 2.23 (3H, s, Ph-CH ), 2.02 (3H, s, pyrimidone-CH ). ESI-MS: m/z 468.4 (M+1) ,
70.5 (M+3), 484.6 (M+Na). C27 OS (467.59).
2
2
S-CH
2
3
3
4
H
N
25 5
4
-((4-(((5-methyl-4-(naphthalen-1-ylmethyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)
methyl)-1H-1,2,3-triazol-1-yl)methyl)benzenesulfonamide (A5b7)
1
Recrystallized from methanol as a white crystal, yield 78.2%, mp:160-162°C. H NMR
400 MHz, DMSO-d6, ppm) δ: 12.56 (1H, s, NH), 8.13 (1H, d, J = 7.72 Hz), 7.86 (1H,
dd, J = 7.28, 1.92 Hz), 7.79 (2H, d, J = 8.28 Hz), 7.73 (1H, d, J = 8.12 Hz), 7.49 (2H, td, J
5.72, 1.64 Hz), 7.29-7.41 (7H, m, aromatic), 5.48 (2H, s, Ph-CH -triazole), 4.37 (2H,
(
=
2
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s, Ph-CH
2
), 4.06 (2H, s, S-CH
2
), 2.04 (3H, s, pyrimidone-CH ). ESI-MS: m/z 533.5
3
(
M+1), 555.3 (M+Na). C H N O S (532.64).
2
6 24 6 3 2
2
-(((1-(4-fluorobenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-5-iodo-6-(naphthalen-1-yl
methyl)pyrimidin-4(3H)-one (A5c1)
1
Recrystallized from methanol as a white crystal, yield 70.3%, mp:131-133°C. H NMR
400 MHz, DMSO-d6, ppm) δ: 13.06 (1H, s, NH), 8.06 (1H, d, J = 8.80 Hz), 7.88 (1H, d,
J = 6.92 Hz), 7.77 (1H, d, J = 8.16 Hz), 7.36-7.52 (4H, m, aromatic), 7.16-7.26 (5H, m,
aromatic), 5.37 (2H, s, Ph-CH -triazole), 4.55(2H, s, Ph-CH ), 3.98 (2H, s, S-CH ).
ESI-MS: m/z 584.1 (M+1), 606.3 (M+Na). C H FIN OS (583.42).
(
2
2
2
2
5
19
5
2
-(((1-benzyl-1H-1,2,3-triazol-4-yl)methyl)thio)-5-iodo-6-(naphthalen-1-ylmethyl)py
rimidin-4(3H)-one (A5c2)
1
Recrystallized from methanol as a light yellow crystal, yield 67.5%, mp: 120-122°C. H
NMR (400 MHz, DMSO-d6, ppm) δ: 13.01 (1H, s, NH), 8.05 (1H, d, J = 7.60 Hz), 7.88
(
1H, dd, J = 7.20, 1.92 Hz), 7.75 (1H, d, J = 8.04 Hz), 7.49 (2H, td, J = 7.16, 1.48 Hz),
.16-7.39 (6H, m, aromatic), 7.14 (2H, dd, J = 7.76, 5.72 Hz), 5.37 (2H, s,
Ph-CH -triazole), 4.54 (2H, s, Ph-CH ), 3.98 (2H, s, S-CH ). ESI-MS: m/z 566.3 (M+1),
88.2 (M+Na). C25 OS (565.43).
7
2
2
2
5
H
20IN
5
5
-iodo-2-(((1-(4-methoxybenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(naphthalen-1-
ylmethyl)pyrimidin-4(3H)-one (A5c3)
1
Recrystallized from methanol as a white crystal, yield 63.4%, mp:192-195°C. H NMR
(
(
400 MHz, DMSO-d6, ppm) δ:13.05 (1H, s, NH), 8.08 (1H, dd, J = 9.00, 1.68 Hz), 7.90
1H, dd, J = 7.04, 2.92 Hz), 7.78 (1H, d, J = 8.08 Hz), 7.52 (2H, td, J = 6.80, 1.76 Hz),
7
.33-7.41 (2H, m, aromatic), 7.18 (1H, s), 7.12 (2H, d, J = 8.60 Hz), 5.28 (2H, s,
1
3
Ph-CH
NMR (100 MHz, DMSO-d6, ppm) δ: 159.6, 143.4, 134.3, 133.8, 132.3, 129.8, 128.9,
28.2, 127.6, 127.4, 126.6, 126.2, 126.0, 124.7, 123.2, 114.5, 55.6, 52.7, 44.1, 24.8.
ESI-MS: m/z 596.3 (M+1), 618.4 (M+Na). C26 S (595.45).
2
-triazole), 4.55 (2H, s, Ph-CH
2
), 3.98 (2H, s, S-CH
2
), 3.73 (3H, s, O-CH
3
). C
1
5 2
H22IN O
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4
-((4-(((5-iodo-4-(naphthalen-1-ylmethyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)met
hyl)-1H-1,2,3-triazol-1-yl)methyl)benzamide (A5c4)
1
Recrystallized from methanol as a light yellow crystal, yield 70.2%, mp:201-203°C. H
NMR (400 MHz, DMSO-d6, ppm) δ:13.085 (1H, s, NH), 8.068 (1H, d, J = 8.00 Hz), 7.98
(
1H, s), 7.85-7.89 (3H, m, aromatic), 7.76 (1H, dd, J = 9.00, 2.16 Hz), 7.52 (2H, td, J =
6
.64, 5.56 Hz), 7.36-7.40 (3H, m, aromatic), 7.20-7.25 (3H, m, aromatic), 5.44 (2H, s,
1
3
Ph-CH
2
-triazole), 4.55 (2H, s, Ph-CH
2
), 4.00 (2H, s, S-CH ). C NMR (100 MHz,
2
DMSO-d6, ppm) δ: 167.8, 143.6, 139.3, 134.5, 134.3, 133.7, 132.3, 128.9, 128.3,
1
6
27.9, 127.6, 127.4, 126.6, 126.2, 125.9, 124.7, 123.7, 52.7, 44.1, 24.7. ESI-MS: m/z
09.2 (M+1), 631.4 (M+Na). C26 S (608.45).
H
21IN
O
6 2
2
-((4-(((5-iodo-4-(naphthalen-1-ylmethyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)met
hyl)-1H-1,2,3-triazol-1-yl)methyl)benzonitrile (A5c5)
1
Recrystallized from methanol as a brown crystal, yield 63.5%, mp:148-151°C. H NMR
(
(
400 MHz, DMSO-d6, ppm) δ:13.06 (1H, s, NH), 8.07 (1H, d, J = 8.24 Hz), 7.86-7.93
2H, m, aromatic), 7.67-7.76 (2H, m, aromatic), 7.47-7.58 (4H, m, aromatic),
7
.19-7.38 (2H, m, aromatic), 7.16 (1H, dd, J = 14.00, 7.68 Hz), 5.61 (2H, s,
Ph-CH -triazole), 4.52 (2H, s, Ph-CH ), 4.00 (2H, s, S-CH ). ESI-MS: m/z 591.3 (M+1) ,
13.2 (M+Na). C26 OS (590.44).
2
2
2
6
H
19IN
6
5
-iodo-2-(((1-(2-methylbenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(naphthalen-1-yl
methyl)pyrimidin-4(3H)-one (A5c6)
1
Recrystallized from methanol as a brown crystal, yield 72.3%, mp:190-192°C. H NMR
400 MHz, DMSO-d6, ppm) δ:13.06 (1H, s, NH), 8.10 (1H, d, J = 7.92 Hz), 7.84 (1H, dd,
(
J = 7.08, 2.12 Hz), 7.71 (1H, d, J = 8.08 Hz), 7.48 (2H, td, J = 5.24, 2.52 Hz), 7.34 (1H, t,
J = 7.40 Hz), 7.21-7.23 (4H, m, aromatic), 7.12 (1H, td, J = 7.32, 2.08 Hz), 6.87 (1H, d, J
=
(
7.44 Hz), 5.42 (2H, s, Ph-CH -triazole), 4.34 (2H, s, Ph-CH ), 3.98 ( 2H, s, S-CH ), 2.25
2
2
2
3H, s, Ph-CH
3
). ESI-MS: m/z 580.3 (M+1) , 602.3 (M+Na). C26
H
22IN
5
OS (579.46).
This article is protected by copyright. All rights reserved.

4
-((4-(((5-iodo-4-(naphthalen-1-ylmethyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)met
hyl)-1H-1,2,3-triazol-1-yl)methyl)benzenesulfonamide (A5c7)
1
Recrystallized from methanol as a light yellow crystal, yield 77.2%, mp:194-197°C. H
NMR (400 MHz, DMSO-d6, ppm) δ: 12.14 (1H, s, NH), 8.12 (1H, d, J = 7.52 Hz), 7.87
(
1H, dd, J = 9.32, 2.28 Hz), 7.78 (2H, d, J = 8.28 Hz), 7.73 (1H, d, J = 8.16 Hz), 7.48 (2H,
td, J = 5.88, 1.80 Hz), 7.29-7.41 (7H, m, aromatic), 5.50 (2H, s, Ph-CH -triazole), 4.43
). ESI-MS: m/z 645.3 (M+1) , 645.3 (M+3) , 667.2
2
(
(
2H, s, Ph-CH
M+Na). C25
2
), 4.03 (2H, s, S-CH
(644.51).
2
H
21IN
6
O S
3 2
4
.1.3 General procedure for the preparation of 5-hydrogen/methyl/iodo-6-(2,6
-
dichlorobenzyl)-2-((1-(arylmethyl)-1H-1,2,3-triazol-4-yl)methylthio)pyrimidin-4(3H
-one(B5a1-B5a7; B5b1-B5b7; B5c1-B5c7).
)
5
-Hydrogen/methyl/iodo-6-(2,6-dichlorobenzyl)-2-((1-(arylmethyl)-1H-1,2,3-triazol-4
-
yl)methylthio)pyrimidin-4(3H)-ones were accomplished efficiently using the
well-established synthetic protocols described in Section 4.1.2.
ethyl 4-(2,6-dichlorophenyl)-3-oxobutanoate (B2a)
1
H NMR (400 MHz, DMSO-d6, ppm) δ: 7.32 (2H, d, J = 8.00 Hz, 3,5-PhH), 7.16 (1H, t, J
=
8.00 Hz, 4-PhH), 4.20-4.25 (4H, m), 3.56 (2H, s,CO-CH
2
-CO), 1.30 (3H, t, J = 8.00 Hz,
CH
3
). C12
H
12Cl
O
2 3
ethyl 4-(2,6-dichlorophenyl)-2-methyl-3-oxobutanoate (B2b)
1
H NMR (400 MHz, DMSO-d6, ppm) δ: 7.32 (2H, d, J = 8.00 Hz, 3,5-PhH), 7.16 (1H, t, J
=
8.00 Hz, 4-PhH), 4.20-4.26 (4H, m), 3.51 (1H, q, J = 8.00 Hz, CO-CH-CO), 1.30 (3H, t,
J = 8.00 Hz, O-CH -CH ), 0.91 (3H, d, J = 8.00 Hz, CH-CH ). C H Cl O
2
3
3
13 14
2 3
6
-(2,6-dichlorobenzyl)-2-thioxo-2,3-dihydropyrimidin-4(1H)-one (B3a)
1
H NMR (400 MHz, DMSO-d6, ppm) δ: 12.47 (1H, s, NH), 12.34 (1H, s, NH), 7.51 (2H,
d, J = 8.00 Hz, 3,5-PhH), 7.34 (1H, t, J = 8.00 Hz, 4-PhH), 5.78 (1H, s, CO-CH), 4.15(2H,
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s, Ph-CH
2
). ESI-MS: m/z 287.3 (M+1), 289.5 (M+3), 291.3 (M+5), 309.5 (M+Na).
C H Cl N OS (285.97).
1
1
8
2 2
6
-(2,6-dichlorobenzyl)-5-methyl-2-thioxo-2,3-dihydropyrimidin-4(1H)-one (B3b)
1
H NMR (400 MHz, DMSO-d6, ppm) δ: 12.47 (1H, s, NH), 12.32 (1H, s, NH), 7.50 (2H,
d, J = 8.00 Hz, 3,5-PhH), 7.34 (1H, t, J = 8.00 Hz, 4-PhH), 4.20(2H, s, Ph-CH ), 1.37 (3H,
2
s, CH
3
). ESI-MS: m/z 301.3 (M+1), 303.4 (M+3), 305.3 (M+5), 323.3 (M+Na).
OS (299.99).
C
12
H
10Cl
N
2 2
6
-(2,6-dichlorobenzyl)-2-(prop-2-yn-1-ylthio)pyrimidin-4(1H)-one(B4a)
1
H NMR (400 MHz, DMSO-d6, ppm) δ: 12.73 (1H, s, NH), 7.51 (2H, d, J = 8.00 Hz,
3
3
,5-PhH), 7.34 (1H, t, J = 8.00 Hz, 4-PhH), 5.78 (1H, s, CO-CH), 4.15(2H, s, Ph-CH
2
),
.88(2H, s, S-CH ), 3.09(1H, s, C≡CH). ESI-MS: m/z 325.3 (M+1), 327.3 (M+3), 329.4
2
(
M+5), 347.2 (M+Na). C H Cl N OS (323.99).
14 10 2 2
6
-(2,6-dichlorobenzyl)-5-methyl-2-(prop-2-yn-1-ylthio)pyrimidin-4(1H)-one (B4b)
1
H NMR (400 MHz, DMSO-d6, ppm) δ: 12.75 (1H, s, NH), 7.50 (2H, d, J = 8.00 Hz,
,5-PhH), 7.34 (1H, t, J = 8.00 Hz, 4-PhH), 4.20 (2H, s, Ph-CH ), 3.91 (2H, s, S-CH ),
3
2
2
3
.08(1H, s, C≡CH), 1.37(3H, s, CH
3
). ESI-MS: m/z 339.3 (M+1), 341.5 (M+3), 343.4
OS (338.00).
(
M+5), 361.5 (M+Na). C15
H
2 2
12Cl N
2
-(((1-(4-acetylbenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(2,6-dichlorobenzyl)pyri
midin-4(3H)-one (B5a1)
1
Recrystallized from methanol as a white crystal, yield 82.3%, mp: 198-200°C. H NMR
400 MHz, DMSO-d6, ppm) δ: 12.63 (1H, s, NH), 7.94 (2H, d, J = 8.2Hz, C ,C -Ph’’-H),
.66 (1H, s, triazole-H), 7.46 (2H, d, J = 8.0 Hz, C ,C -Ph’-H), 7.38 (2H, d, J = 8.2 Hz,
(
3
5
7
C
2
6
3
,C
4
-Ph’-H), 7.30 (1H, t, J = 7.8 Hz, C
), 4.35 (2H, s, S-CH ), 4.15 (2H, s, Ph-CH
100 MHz, DMSO-d6, ppm) δ: 197.93, 172.46, 164.11, 143.84, 141.45, 136.94,
4
-Ph’’-H), 5.74 (1H , s, pyrimidin-H), 5.63 (2H , s,
1
3
triazole-CH
2
2
2
), 2.56 (3H, s, CO-CH ). C NMR
3
(
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1
35.99, 133.56, 130.16, 129.12, 128.93, 128.41, 124.02, 52.80, 38.39, 24.85, 21.51.
ESI-MS: m/z 500.2 (M+1), 522.2 (M+Na). C H Cl N O S (499.06).
2
3
19
2 5 2
6
-(2,6-dichlorobenzyl)-2-(((1-(4-fluorobenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)pyri
midin-4(3H)-one (B5a2)
1
Recrystallized from methanol as a white crystal, yield 84.5%,mp: 189-191°C. H NMR
400 MHz, DMSO-d6, ppm) δ:12.62 (1H, s, NH), 7.76 (1H, s, triazole-H), 7.46 (2H, d, J
8.0Hz, C ,C -Ph’’-H), 7.29-7.37 (3H, m, aromatic), 7.18 (1H, t, J = 7.8 Hz), 5.74 (1H, s,
pyrimidone-H), 5.53 (2H, s, triazole-CH
NMR (100 MHz, DMSO-d6, ppm) δ: 163.57, 161.14, 143.84, 136.01, 133.61, 132.63,
(
=
3
5
1
3
2
), 4.34 (2H, s, S-CH
2
), 4.14 (2H, s, Ph-CH ). C
2
1
3
30.61 (d, J = 8.0 Hz, C
3
,C
5
-Ph’’), 130.11, 128.90, 123.71, 116.15, 115.93, 52.49,
FN OS (475.04).
8.45, 24.89.ESI-MS: m/z476.2 (M+1), 498.3 (M+Na). C21
H
16Cl
2
5
2
-(((1-benzyl-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(2,6-dichlorobenzyl)pyrimidin-4(3
H)-one (B5a3)
1
Recrystallized from methanol as a white crystal, yield 80.5%,mp: 218-221°C. H NMR
400 MHz, DMSO-d6, ppm) δ:12.64 (1H, s, NH), 7.74 (1H, s, triazole-H), 7.45 (2H, d, J
8.0Hz, C ,C -Ph’’-H), 7.26-7.39(6H, m, aromatic), 5.75 (1H, s, pyrimidone-H), 5.53
2H, s, triazole-CH
DMSO-d6, ppm) δ: 143.72, 136.41, 136.01, 133.59, 130.14, 129.21, 128.92, 128.60,
(
=
3
5
1
3
(
2
), 4.33 (2H, s, S-CH
2
), 4.15 ( 2H, s, Ph-CH ). C NMR (100 MHz,
2
1
28.31, 123.74, 53.26, 38.42, 24.86. ESI-MS: m/z 458.4 (M+1), 480.3 (M+Na).
OS (457.05).
C
21
H
2 5
17Cl N
4
3
-((4-(((4-(2,6-dichlorobenzyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)-1H-1,2,
-triazol-1-yl)methyl)benzamide (B5a4)
1
Recrystallized from methanol as a white crystal, yield 79.6%, mp: 248-250°C. H NMR
(
400 MHz, DMSO-d6, ppm) δ:12.65 (1H, s, NH), 7.97 (1H, s, triazole-H), 7.86 (2H , d, J
8.1Hz, C ,C -Ph’-H), 7.78 (1H , s), 7.45 (2H, d, J = 8.0Hz, C ,C -Ph’’-H), 7.28-7.38 (4H,
=
3
5
3
5
m, aromatic), 5.77 (1H, s, pyrimidone-H), 5.60 (2H, s, triazole-CH
2
), 4.35 (2H, s,
1
3
S-CH
2
), 4.16 (2H, s, Ph-CH
2
). C NMR (100 MHz, DMSO-d6, ppm) δ: 167.89, 164.39,
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1
1
62.76, 143.81, 139.41, 136.01, 134.56, 133.61, 130.10, 128.90, 128.38, 128.05,
23.93, 52.87, 38.42, 24.87. ESI-MS: m/z 501.3 (M+1), 523.4 (M+Na). C H Cl N O S
2
2
18
2 6 2
(
500.06).
6
-(2,6-dichlorobenzyl)-2-(((1-(4-methoxybenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)p
yrimidin-4(3H)-one (B5a5)
1
Recrystallized from methanol as a white crystal, yield 84.1%,mp: 174-176°C. H NMR
400 MHz, DMSO-d6, ppm) δ:12.57 (1H, s, NH), 7.70 (1H , s, triazole-H),7.46 (2H, d, J
7.9Hz, C ,C -Ph’’-H), 7.28 (1H, t, J = 7.8 Hz, C -Ph’’-H), 7.24 (2H , d, J = 7.6Hz,
C ,C -Ph’-H), 6.91 (2H, d, J = 7.7Hz, C ,C -Ph’-H), 5.73 (1H, s, pyrimidone-H), 5.44 (2H,
(
=
3
5
4
2
6
3
5
1
3
s, triazole-CH
2
), 4.32 (2H, s, S-CH
2
), 4.11 ( 2H, s, Ph-CH
2
), 3.73 ( 3H, s, O-CH ). C NMR
3
(
100 MHz, DMSO-d6, ppm) δ: 159.62, 136.02, 133.62, 130.13, 130.02, 128.90,
1
14.57, 55.59, 52.84, 38.45, 24.23. ESI-MS: m/z 488.3 (M+1), 510.3 (M+Na).
S (487.06).
C
22
H
2
19Cl N
O
5 2
4
3
-((4-(((4-(2,6-dichlorobenzyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)-1H-1,2,
-triazol-1-yl)methyl)benzonitrile (B5a6)
1
Recrystallized from methanol as a white crystal, yield 76.8%,mp: 215-217°C. H NMR
(
400 MHz, DMSO-d6, ppm) δ:12.60 (1H, s, NH), 7.84 (2H, d, J = 8.2Hz, C
3
,C
-Ph’-H), 7.40 (2H, d, J = 8.1Hz,
-Ph’’-H), 5.75 (1H , s, pyrimidone-H), 5.66 (2H,
5
-Ph’’-H),
7
.82 (1H , s, triazole-H), 7.46 (2H , d, J = 8.0Hz, C
,C -Ph’-H), 7.28 (1H, t, J = 7.9 Hz, C
), 4.35 (2H, s, S-CH ), 4.15 (2H, s, Ph-CH
DMSO-d6, ppm) δ: 170.79, 143,94, 141.91, 135.99, 133.58, 133.19, 130.14, 129.04,
28.92, 124.20, 118.98, 111.40, 52.62, 38.42, 24.84.ESI-MS: m/z 483.2 (M+1), 505.2
M+Na)C22 OS (482.05).
3
,C
5
C
2
6
4
1
3
s, triazole-CH
2
2
2
). C NMR (100 MHz,
1
(
H
16Cl
N
2 6
4
3
-((4-(((4-(2,6-dichlorobenzyl)-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)-1H-1,2,
-triazol-1-yl)methyl)benzenesulfonamide (B5a7)
1
Recrystallized from methanol as a white crystal, yield 83.2%, mp: 229-231°C. H NMR
(
400 MHz, DMSO-d6, ppm) δ:12.67 (1H, s, NH), 7.81 (2H , d, J = 8.2Hz, C
3
5
,C -Ph’-H),
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7
.80 (1H, s, triazole-H), 7.34-7.48 (6H , m, aromatic), 7.28 (1H, t, J = 8.0 Hz, C
4
-Ph’’-H),
5
.76 (1H, s, pyrimidone-H), 5.62 (2H, s, triazole-CH ), 4.34 (2H, s, S-CH ), 4.16 (2H, s,
2
2
13
Ph-CH ). C NMR (100 MHz, DMSO-d6, ppm) δ:163.25, 144.32, 143.84, 140.17,
2
1
36.01, 133.61, 130.13, 128.92, 128.75, 126.58, 124.00, 52.66, 38.43, 24.86. ESI-MS:
m/z537.2 (M+1), 559.2 (M+Na). C21 (536.03).
H
18Cl
2
N
6
O S
3 2
2
-(((1-(4-acetylbenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(2,6-dichlorobenzyl)-5-m
ethylpyrimidin-4(3H)-one(B5b1)
1
Recrystallized from methanol as a white crystal, yield 85.3%,mp: 207-209°C. H NMR
400 MHz, DMSO-d6, ppm) δ:12.52 (1H, s, NH), 7.95 (2H, d, J = 8.2Hz, C ,C -Ph’’-H),
.33-7.37 (5H , m, aromatic), 7.14 (1H, t, J = 7.8 Hz, C -Ph’’-H), 5.59 (2H, s,
triazole-CH ), 4.14 (2H, s, S-CH ), 4.08 ( 2H, s, Ph-CH ), 2.57 (3H, s, COCH ), 2.06 (3H,
(
3
5
7
4
2
2
2
3
1
3
s, CH ). C NMR (100 MHz, DMSO-d6, ppm) δ: 197.95, 163.90, 158.73, 141.37,
3
1
3
36.96, 136.02, 135.90, 135.05, 129.45, 129.12, 128.45, 128.39, 123.39, 52.80,
5.62, 27.23, 24.24, 10.59. ESI-MS: m/z514.4 (M+1), 536.2 (M+Na). C H Cl N O S
2
4
21
2 5 2
(
513.08).
6
-(2,6-dichlorobenzyl)-2-(((1-(4-fluorobenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-5-m
ethylpyrimidin-4(3H)-one (B5b2)
1
Recrystallized from methanol as a white crystal, yield 82.4%, mp: 224-226°C. H NMR
(
400 MHz, DMSO-d6, ppm) δ:12.68 (1H, s, NH), 7.32-7.35(5H, m, aromatic), 7.15-7.24
(
3H, m, aromatic), 5.48 (2H, s, triazole-CH ), 4.17 (2H, s, S-CH ), 4.02 (2H, s, Ph-CH ),
2
2
2
1
3
2
1
1
.06 (3H, s, CH
36.01, 135.05, 132.61, 132.58, 130.63 (d, J = 9.0 Hz, C
23.05, 116.16, 115.94, 52.45, 35.63, 24.25, 10.58. ESI-MS: m/z 490.3 (M+1), 512.4
FN OS (489.06).
3
). C NMR (100 MHz, DMSO-d6, ppm) δ: 163.58, 161.15, 143.95,
3
5
,C -Ph’’), 129.42, 128.38,
(
M+Na). C22
H
18Cl
2
5
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2
-(((1-benzyl-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(2,6-dichlorobenzyl)-5-methylpyri
midin-4(3H)-one (B5b3)
1
Recrystallized from methanol as a light yellow crystal, yield 76.4%,mp: 240-242°C. H
NMR (400 MHz, DMSO-d6, ppm) δ:12.59 (1H, s, NH), 7.35-7.40 (4H , m,
aromatic),7.31 (2H , d, J = 8.0Hz, C ,C -Ph’-H), 7.24 (2H, d, J = 6.8Hz, C ,C -Ph’-H),
3
5
2
6
7
.26-7.39 (6H, m, aromatic), 7.14 (1H, t, J = 7.9 Hz, C
4
-Ph’’-H), 5.48 (2H, s,
1
3
triazole-CH ), 4.17 (2H, s, S-CH ), 4.04 (2H, s, Ph-CH
2
2
2
), 2.06 (3H, s, CH ). C NMR (100
3
MHz, DMSO-d6, ppm) δ: 170.78, 163.65, 144.26, 136.32, 136.03, 135.07, 129.42,
1
4
29.20, 128.64, 128.36, 123.11, 53.27, 35.64, 24.26, 10.59. ESI-MS: m/z 472.3 (M+1),
94.3 (M+Na). C22 OS (471.07).
H
19Cl
N
2 5
4
-((4-(((4-(2,6-dichlorobenzyl)-5-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)thio)meth
yl)-1H-1,2,3-triazol-1-yl)methyl)benzamide (B5b4)
1
Recrystallized from methanol as a white crystal, yield 78.7%,mp: 267-269°C. H NMR
400 MHz, DMSO-d6, ppm) δ:12.52 (1H, s, NH), 7.97 (1H, s, triazole-H),7.86 (2H, d, J =
.1Hz, C ,C -Ph’-H), 7.39 (1H , s), 7.34 (2H, d, J = 6.6Hz, C ,C -Ph’’-H), 7.28-7.34 (3H,
(
8
3
5
3
5
m, aromatic), 7.13 (1H, t, J = 8.0 Hz, C
4
-Ph’’-H), 5.54 (2H, s, triazole-CH
2
), 4.17 (2H, s,
1
3
S-CH ), 4.02 (2H, s, Ph-CH ), 2.05 (3H, s, CH
2
2
3
). C NMR (100 MHz, DMSO-d6, ppm) δ:
1
1
C
67.85, 144.02, 139.36, 136.01, 135.05, 134.54, 129.44, 128.39, 128.37, 128.09,
23.29, 52.83, 35.63, 24.22, 10.60. ESI-MS: m/z515.3 (M+1), 537.2 (M+Na).
23
H
2
20Cl N
6
O
2
S (514.07).
6
-(2,6-dichlorobenzyl)-2-(((1-(4-methoxybenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-5
-
methylpyrimidin-4(3H)-one (B5b5)
1
Recrystallized from methanol as a yellow crystal, yield 80.2%,mp: 184-186°C. H NMR
400 MHz, DMSO-d6, ppm) δ:12.62 (1H, s, NH), 7.33 (2H, d, J = 8.0Hz, C ,C -Ph’’-H),
.28 (1H, s, triazole-H), 7.22 (2H, d, J = 8.6Hz, C ,C -Ph’-H), 7.15 (1H, t, J = 7.8 Hz,
(
3
5
7
C
2
6
4
-Ph’’-H), 6.92 (2H, dd, J = 6.7, 1.8Hz, C
3
,C
5
-Ph’-H), 5.39 (2H , s, triazole-CH
2
), 4.17
1
3
(
2H, s, S-CH
2
), 4.01 ( 2H, s, Ph-CH ), 3.74 (3H, s, OCH
2
3
), 2.06 (3H, s, CH
3
). C NMR
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(
100 MHz, DMSO-d6, ppm) δ: 159.62, 143.81, 136.01, 135.05, 130.00, 129.45,
28.40, 128.22, 122.76, 114.58, 55.61, 52.80, 35.63, 24.23, 10.59. ESI-MS: m/z502.3
M+1), 524.3 (M+Na) C23 S (501.08).
1
(
H
21Cl
2
N O
5 2
4
-((4-(((4-(2,6-dichlorobenzyl)-5-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)thio)meth
yl)-1H-1,2,3-triazol-1-yl)methyl)benzonitrile (B5b6)
1
Recrystallized from methanol as a light yellow crystal, yield 82.7%, mp: 223-225°C. H
NMR (400 MHz, DMSO-d6, ppm) δ: 12.59 (1H, s, NH), 7.86 (2H, d, J = 8.8Hz,
C
3
,C
5
-Ph’’-H), 7.40-7.41 (3H , m, aromatic), 7.33 (2H, d, J = 8.0Hz, C
2
6
,C -Ph’-H), 7.14
(
1H, t, J = 7.9 Hz, C -Ph’’-H), 5.62 (2H, s, triazole-CH ), 4.17 (2H, s, S-CH ), 4.04(2H, s,
4
2
2
1
3
Ph-CH
2
), 2.06 (3H, s, CH ). C NMR (100 MHz, DMSO-d6, ppm) δ: 163.70, 144.17,
3
1
5
41.80, 136.01, 135.05, 133.18, 129.41, 129.10, 128.37, 123.57, 118.98, 111.43,
2.63, 35.63, 24.26, 10.59. ESI-MS: m/z 497.4 (M+1), 519.2 (M+Na) C H Cl N OS
2
3
18
2 6
(
496.06).
4
-((4-(((4-(2,6-dichlorobenzyl)-5-methyl-6-oxo-1,6-dihydropyrimidin-2-yl)thio)meth
yl)-1H-1,2,3-triazol-1-yl)methyl)benzenesulfonamide (B5b7)
1
Recrystallized from methanol as a light yellow crystal, yield 78.3%,mp: 243-245°C. H
NMR (400 MHz, DMSO-d6, ppm) δ: 12.64 (1H, s, NH), 7.82 (2H, d, J = 8.2Hz,
C
3
,C
5
-Ph’-H), 7.38-7.42 (5H, m, aromatic), 7.33 (2H, d, J = 8.0Hz, C
2
,C
), 4.18 (2H, s, S-CH
). C NMR (100 MHz, DMSO-d6, ppm) δ: 163.25, 144.34,
6
-Ph’-H), 7.16
(
1H, t, J = 8.0 Hz, C
4
-Ph’’-H), 5.58 (2H, s, triazole-CH
2
2
), 4.03 (2H, s,
1
3
Ph-CH
2
), 2.06 (3H, s, CH
3
1
3
44.09, 140.09, 136.03, 135.05, 129.43, 128.78, 128.39, 126.58, 123.37, 52.64,
5.64, 24.27, 10.60. ESI-MS: m/z551.3 (M+1), 573.1 (M+Na). C H Cl N O S
2
2
20
2 6 3 2
(
550.04).
2
-(((1-(4-acetylbenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(2,6-dichlorobenzyl)-5-io
dopyrimidin-4(3H)-one (B5c1)
1
Recrystallized from methanol as a white crystal, yield 84.2%,mp: 213-215°C. H NMR
400 MHz, DMSO-d6, ppm) δ: 13.11 (1H, s, NH), 7.95 (2H, d, J = 8.2Hz, C ,C -Ph’’-H),
(
3
5
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7
.45 (1H, s, triazole-H), 7.36 (2H, d, J = 8.2 Hz, C
2
6
,C -Ph’-H), 7.32 (2H, d, J = 8.0 Hz,
C ,C -Ph’-H), 7.16 (1H, t, J = 7.7 Hz, C -Ph’’-H), 5.59 (2H, s, triazole-CH ), 4.29 (2H, s,
3
4
4
2
1
3
S-CH
2
), 4.03 ( 2H, s, Ph-CH
2
), 2.57 (3H, s, CO-CH ). C NMR (100 MHz, DMSO-d6,
3
ppm) δ: 197.95, 164.51, 143.62, 141.34, 136.97, 136.03, 135.93, 134.75, 129.73,
1
6
29.12, 128.48, 128.37, 123.56, 52.82, 42.71, 27.25, 24.66. ESI-MS: m/z 626.1 (M+1),
48.2 (M+Na). C23 IN S (624.96).
H
18Cl
2
O
5 2
6
-(2,6-dichlorobenzyl)-2-(((1-(4-fluorobenzyl)-1H-1,2,3-triazol-4-yl)methyl)thio)-5-io
dopyrimidin-4(3H)-one (B5c2)
1
Recrystallized from methanol as a white crystal, yield 82.3%,mp: 221-224°C. H NMR
400 MHz, DMSO-d6, ppm)δ: 12.87 (1H, s, NH), 7.42 (1H, s, triazole-H), 7.31-7.36 (4H,
m, aromatic), 7.16-7.24 (3H, m, aromatic), 5.48 (2H, s, triazole-CH ), 4.28 (2H, s,
(
2
1
3
S-CH
2
), 3.96 (2H, s, Ph-CH
63.58, 143.85, 136.00, 134.96, 133.59, 130.66 (d, J = 9.0 Hz, C
28.91, 128.32, 123.16, 116.16, 115.94, 52.47, 42.73, 24.70. ESI-MS: m/z 602.2
FIN OS (600.94).
2
). C NMR (100 MHz, DMSO-d6, ppm) δ: 172.45, 164.15,
1
1
3
5
,C -Ph’’), 130.13,
(
M+1), 624.2 (M+Na). C21
H
15Cl
2
5
2
-(((1-benzyl-1H-1,2,3-triazol-4-yl)methyl)thio)-6-(2,6-dichlorobenzyl)-5-iodopyrimi
din-4(3H)-one (B5c3)
1
Recrystallized from methanol as a white crystal, yield 86.2%,mp: 215-217°C. H NMR
(
400 MHz, DMSO-d6, ppm) δ: 13.12 (1H, s, NH), 7.33-7.36 (4H, m, aromatic), 7.28
2H , d, J = 8.5Hz, C ,C -Ph’-H), 7.26 (2H , d, J = 6.6Hz, C ,C -Ph’-H), 7.13 (1H, t, J = 7.7
(
3
5
2
6
1
3
Hz, C
4
-Ph’’-H), 5.49 (2H, s, triazole-CH
2
), 4.29 (2H, s, S-CH
2
), 4.02 (2H, s, Ph-CH ). C
2
NMR (100 MHz, DMSO-d6, ppm) δ: 143.47, 136.32, 136.04, 134.77, 129.70, 129.21,
28.63, 128.38, 128.36, 123.25, 53.30, 42.71, 24.69. ESI-MS: m/z584.0 (M+1), 606.1
M+Na). C21 IN OS (582.95).
1
(
H
16Cl
2
5
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4
1
-((4-(((4-(2,6-dichlorobenzyl)-5-iodo-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)-
H-1,2,3-triazol-1-yl)methyl)benzamide (B5c4)
1
Recrystallized from methanol as a white crystal, yield 79.3%,mp: 248-250°C. H NMR
(
400 MHz, DMSO-d6, ppm) δ: 13.12 (1H, s, NH), 7.96 (1H, s, triazole-H),7.86 (2H, d, J
=
5
8.2Hz, C ,C -Ph’-H), 7.30-7.44 (6H, m, aromatic), 7.15 (1H, t, J = 7.8 Hz, C -Ph’’-H),
3 5 4
1
3
.55 (2H , s, triazole-CH
2
), 4.29 (2H, s, S-CH
2
), 3.99 ( 2H, s, Ph-CH ). C NMR (100
2
MHz, DMSO-d6, ppm) δ: 167.86, 143.60, 139.33, 136.03, 134.79, 134.57, 129.70,
28.37, 128.12, 123.46, 52.86, 42.72, 24.66. ESI-MS: m/z627.1 (M+1), 649.2 (M+Na).
C H Cl IN O S (625.96).
1
2
2
17
2
6 2
6
-(2,6-dichlorobenzyl)-5-iodo-2-(((1-(4-methoxybenzyl)-1H-1,2,3-triazol-4-yl)methyl
)thio)pyrimidin-4(3H)-one(B5c5)
1
Recrystallized from methanol as a light yellow crystal, yield 73.6%,mp: 206-209°C. H
NMR (400 MHz, DMSO-d6, ppm) δ: 13.14 (1H, s, NH), 7.36 (1H , s, triazole-H), 7.46
(
2H, d, J = 8.0Hz, C
3
,C
5
-Ph’’-H), 7.24 (2H, d, J = 8.5Hz, C
2
6
,C -Ph’-H), 7.18 (1H, t, J = 7.9
Hz, C -Ph’’-H), 6.93 (2H, d, J = 8.6Hz, C ,C -Ph’-H), 5.40 (2H, s, triazole-CH ), 4.29 (2H,
4
3
5
2
1
3
s, S-CH
2
), 3.97 (2H, s, Ph-CH
2
), 3.74 (3H, s, O-CH ). C NMR (100 MHz, DMSO-d6,
3
ppm) δ: 164.40, 159.62, 143.35, 136.03, 134.76, 130.04, 129.74, 128.37, 128.21,
22.94, 114.57, 55.62, 52.83, 42.70, 24.65. ESI-MS: m/z614.1 (M+1), 636.1 (M+Na).
C H Cl IN O S (612.96).
1
2
2
18
2
5 2
4
1
-((4-(((4-(2,6-dichlorobenzyl)-5-iodo-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)-
H-1,2,3-triazol-1-yl)methyl)benzonitrile (B5c6)
1
Recrystallized from methanol as a white crystal, yield 76.6%,mp: 226-229°C. H NMR
(
400 MHz, DMSO-d6, ppm) δ: 13.16 (1H, s, NH), 7.87 (2H, d, J = 8.2Hz, C
.48 (1H, s, triazole-H), 7.41 (2H, d, J = 8.2Hz, C ,C -Ph’-H), 7.32 (2H, d, J = 8.0Hz,
C ,C -Ph’-H), 7.16 (1H, t, J = 7.6 Hz, C -Ph’’-H), 5.63 (2H, s, triazole-CH ), 4.30 (2H, s,
3
,C -Ph’’-H),
5
7
3
5
2
6
4
2
1
3
S-CH
2
), 4.15 (2H, s, Ph-CH ). C NMR (100 MHz, DMSO-d6, ppm) δ: 164.46, 143.70,
2
1
41.76, 136.04, 134.74, 133.18, 129.69, 129.04, 128.34, 123.73, 118.96, 111.46,
This article is protected by copyright. All rights reserved.

5
2.66, 42.71, 24.69. ESI-MS: m/z609.1 (M+1), 631.2 (M+Na). C22
H
15Cl
2
IN OS
6
(
607.94).
4
1
-((4-(((4-(2,6-dichlorobenzyl)-5-iodo-6-oxo-1,6-dihydropyrimidin-2-yl)thio)methyl)-
H-1,2,3-triazol-1-yl)methyl)benzenesulfonamide (B5c7)
1
Recrystallized from methanol as a white crystal, yield 74.3%,mp: 221-223°C. H NMR
(
400 MHz, DMSO-d6, ppm) δ: 13.14 (1H, s, NH), 7.82 (2H, d, J = 8.1Hz, C
.46 (1H, s, triazole-H), 7.41 (2H, d, J = 8.1Hz, C ,C -Ph’’-H), 7.32-7.36 (4H, m,
aromatic), 7.15 (1H, t, J = 8.0 Hz, C -Ph’’-H), 5.58 (2H, s, triazole-CH ), 4.29 (2H, s,
S-CH ), 3.99 (2H, s, Ph-CH ). C NMR (100 MHz, DMSO-d6, ppm) δ:163.24, 144.36,
3
,C -Ph’-H),
5
7
3
5
4
2
1
3
2
2
1
2
40.04, 136.04, 134.77, 129.72, 128.82, 128.36, 126.57, 107.42, 52.67, 42.73,
4.67.ESI-MS: m/z663.1 (M+1), 685.1 (M+Na). C21
H
17Cl
2
IN
6
O
3
S (661.92).
2
4
.2. In vitro anti-HIV assay
Evaluation of the antiviral activity of the compounds against HIV-1 strain IIIB
and HIV-2 strain (ROD) in MT-4 cells was performed using the MTT assay as
previously described [15,16]. Stock solutions (10×final concentration) of test
compounds were added in 25µL volumes to two series of triplicate wells so as to
allow simultaneous evaluation of their effects on mock- and HIV-infected cells at the
beginning of each experiment. Serial fivefold dilutions of test compounds were made
directly in flat-bottomed 96 wells microtiter trays using a Biomek 3000 robot
(
Beckman instruments, Fuller-ton, CA). Untreated control HIV- and mock-infected
cell samples were included for each sample.
HIV-1 (IIIB) or HIV-2 (ROD) [17] stock (50µL) at 100–300 CCID50 (cell culture
infectious dose) or culture medium was added to either the infected or
mock-infected wells of the microtiter tray. Mock-infected cells were used to evaluate
the effect of test com-pound on uninfected cells in order to assess the cytotoxicity of
the test compound. Exponentially growing MT-4 cells were cen-trifuged for 5 min at
1
000 rpm and the supernatant was discarded. The MT-4 cells were resuspended at
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5
6
×10 cells/mL, and 50-µL volumes were transferred to the microtiter tray wells. Five
days after infection, the viability of mock- and HIV-infected cells was examined
spectrophotometrically by the MTT assay.
The MTT assay is based on the reduction of yellow colored
3
-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) (Acros Organics,
Geel, Belgium) by mitochondrial dehydro-genase of metabolically active cells to a
blue-purple formazan that can be measured spectrophotometrically. The
absorbances were read in an eight-channel computer-controlled photometer
(
Multi-scan Ascent Reader, Labsystems, Helsinki, Finland), at two wave-lengths (540
and 690 nm). All data were calculated using the median OD (optical density) value of
tree wells. The 50% cytotoxic concentration (CC50) was defined as the concentration
of the test compound that reduced the absorbance (OD540) of the mock-in-fected
control sample by 50%. The concentration achieving 50% protection from the
cytopathic effect of the virus in infected cells was defined as the 50% effective
concentration (EC50).
Acknowledgment
The financial support from the Key Project of National Natural Science Foundation of
China (NNSFC) for International Cooperation (No. 81420108027), National Natural
Science Foundation of China (No.81102320, 81273354), Research Fund for the
Doctoral Program of Higher Education of China (No. 20110131130005,
2
0110131120037), The Science and Technology Development Project of Shandong
Province (No. 2014GSF118012), China Postdoctoral Science Foundation Funded
Special Project (No. 2012T50584) and KU Leuven (GOA 10/014) is gratefully
acknowledged.
Conflict of Interest
The authors have declared no conflict of interest.
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