Synthesis and biological evaluation of novel 1,2,3-triazolonucleotides

Article abstract of DOI:10.1002/ardp.201200421

A general procedure for the preparation of 1,2,3-triazole analogs of nucleosides from diethyl 2-azidoethoxymethyl- and 2-azidoethoxyethylphosphonates was elaborated. The application of microwave irradiation shortened the reaction time to 10 min in comparison to ca. 48 h when 1,3-dipolar cycloadditions were performed under standard conditions. All compounds were evaluated in vitro for inhibitory activity against a broad variety of DNA and RNA viruses. None of the compounds were antivirally active at subtoxic concentrations. Compound 17k exhibited moderate inhibitory effects on the proliferation of human T-lymphocyte cells (IC₅₀ = 64 μM for CEM). The inhibitory activity of cell proliferation for HEL cells as well as L1210, CEM, and HeLa cells of several 1,2,3-triazole analogs of nucleotides was evaluated. The 1,2,3-triazole derivatives appeared to be cytostatic (with IC₅₀ values reaching the middle to higher micromolar range: 60-250 μM). Copyright

Full text of DOI:10.1002/ardp.201200421

Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
1
Full Paper
Synthesis and Biological Evaluation of Novel
1,2,3-Triazolonucleotides
1
Iwona E. Głowacka , Jan Balzarini , and Andrzej E. Wroblewski
2
1
´
1
´ ´ ´ ´
Faculty of Pharmacy, Bioorganic Chemistry Laboratory, Medical University of Łodz, Łodz, Poland
2 Rega Institute for Medical Research, KU Leuven, Leuven, Belgium
A general procedure for the preparation of 1,2,3-triazole analogs of nucleosides from diethyl 2-
azidoethoxymethyl- and 2-azidoethoxyethylphosphonates was elaborated. The application of
microwave irradiation shortened the reaction time to 10 min in comparison to ca. 48 h when 1,3-
dipolar cycloadditions were performed under standard conditions. All compounds were evaluated
in vitro for inhibitory activity against a broad variety of DNA and RNA viruses. None of the compounds
were antivirally active at subtoxic concentrations. Compound 17k exhibited moderate inhibitory
effects on the proliferation of human T-lymphocyte cells (IC50 ¼ 64 mM for CEM).
Keywords: Antiviral / Aziodophosphonates / Cycloaddition / Cytostatic / 1,2,3-Triazoles
Received: November 5, 2012; Accepted: December 21, 2012
DOI 10.1002/ardp.201200421
Introduction
conformational flexibility of the nucleoside molecule and
in lowering electrostatic repulsion [16–18].
Phosphonate nucleoside analogs are recognized as com-
pounds possessing a broad spectrum of antiviral and cyto-
static activities [1, 2]. Antiviral medications used so far
have several limitations including long-term side-effects.
Furthermore, the possibility of mutation of various viruses
brings an important problem in the treatment of viral infec-
tions. For this reason a search for new antiviral compounds
has been underway for many years. Many structural modifi-
cations have been described for both heterocyclic base and
sugar moieties. For example, nucleoside analogs in which the
furanose ring has been replaced by heterocyclic rings such as
isoxazole 1 [3, 4], isoxazoline 2 [5, 6], cis- and trans-isoxazoli-
dine 3 [7–9], and 1,2,3-triazole 4–10 [10–15] have been
reported and some of them showed interesting antiviral
and/or anticancer activities (Fig. 1).
On the other hand, several nucleoside analogs composed of
nucleobases and 1,2,3-triazoles have been described in the
literature (Fig. 2) [19–22].
The interest in incorporation of an 1,2,3-triazole ring into
various classes of compounds is stimulated by recent discov-
eries of their biological activity and potential applications as
antibacterial [23–25], antifungal [26–28], anticancer [21, 29–
33], and antiviral [10, 34, 35] agents. Furthermore, some
substituted 1,2,3-triazoles are also used as agrochemicals,
dyes, photostabilizers, and corrosion inhibitors [36–39].
The straightforward synthesis of 1,2,3-triazoles is based on
the Huisgen cycloaddition of azides and alkynes. However,
under thermal conditions the reaction with terminal alkynes
gives a mixture of 1,4- and 1,5-disubstituted regioisomers
[40, 41]. The application of copper(I) salts as catalyst led to
the formation of the 1,4-disubstituted isomers, regiospecifically
[42, 43].
The b-N-glycosidic bonds found in the natural nucleosides
significantly contribute to the typical conformational pref-
erences of the nucleosides. Insertion of a methylene group
between a nucleobase and a sugar ring results in higher
It has been firmly established that nucleoside analogs need
to be phosphorylated intracellularly into the respective tri-
phosphates (active antimetabolites) to display antiviral
activity [44].
On the other hand, the nucleotide analogs (adefovir, teno-
fovir, cidofovir) containing a stable isopolar phosphono-
methyl ether (P–CH2–O) moiety instead of the nucleotide
phosphate ester group (P–O–C) are resistant to enzymatic
Correspondence: Dr. Iwona E. Głowacka, Faculty of Pharmacy,
´
Bioorganic Chemistry Laboratory, Medical University of Łodz,
´
´
´
´
Muszynskiego 1, 90-151 Łodz, Poland.
E-mail: iwona.glowacka@umed.lodz.pl
Fax: þ48 42 678 83 98
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

2
I. E. Głowacka et al.
Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
O
N
O
O
N
R'
O
Bz
O
NH
NH
N
OH
BocHN
R
NH
t-BuO
O
N
O
H3C
N
N
N
O
O
O
H3C
3
2
1
a: R = CH2C6H5, R' = CH3
b: R = CH(CH3)2, R' = H
O
OCH3
SCH2C6H5
I
NH
N
N
N
N
N
N
N
N
N
O
N
N
N
O
N
O
N
N
N
N
N
HO
HO
HO
4
6
5
H2N
O
N
N
NH
N
N
B
N
N
O
OH
B
N
N
N
N
(HO)2(O)P
N
N
N
N
N
HO
(HO)2(O)P
N
N
9
7
8
10
a: B = Adenine
b: B = Thymine
a: B = Adenine
b: B = Thymine
c: B = 6-Azauracil
d: B = Uracil
Figure 1. Nucleoside and nucleotide analogs with heterocyclic rings.
degradation and the problem of intracellular phosphoryl-
ation necessary for nucleoside activation is avoided [45,
(Fig. 3) containing
(P–CH2CH2–O).
a
phosphonoethyl ether moiety
´
46]. Furthermore, in 2009, Hockova and co-workers reported
In continuation of our research program on the chemistry
of 1,2,3-triazoles [49–52] and 1,2,3-triazole nucleotide analogs
[53], new series of compounds of general formula 17–20 were
synthesis and antimalarial activity of several acyclic nucleo-
tide phosphonates PEEG, PEEHx [47], and their analogs [48]
Nucleobase
O
N
O
N
NH
NH
Nucleobase
N
N
N
N
N
N
Nucleobase
O
O
N
O
O
HO
HO
Me
O
O
N
N
N
(EtO)2(O)P
R2
R1
N
OH
Nucleobase
N
R1 = OH, R2 = H
R1 = R2 = SEt
N
Figure 2. Nucleoside analogs having natural nucleobases connected with the 1,2,3-triazole ring by a methylene linker.
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
Synthesis and Biological Evaluation of 1,2,3-Triazolonucleotides
3
O
O
N
N
NH
NH
X
N
N
N
R
(HO)2(O)P
N
R
(HO)2(O)P
O
O
R = NH2, PEEG
R = H, NH2
X = CH3, CH2CH3, CH2C6H5
R = H,
PEEHx
Figure 3. Structures of PEEG, PEEHx and
their analogs.
B
m
N
(EtO)2(O)P
N3
(EtO)2(O)P
N
N
(EtO)2(O)P
Cl
O
O
O
n
n
n
_
n = 0, m = 1 17a l
n = 0 11
n = 1 12
n = 0 13
n = 1 14
_
n = 1, m = 1 18a l
_
n = 0, m = 0 19a c
_
n = 1, m = 0 20a c
B - nucleobases or their mimetics
Scheme 1. Retrosynthetic analysis of 1,2,3-triazoles.
designed which incorporate the known structural features of
active phosphonate nucleoside analogs such as P–CH2–O and
P–CH2CH2–O fragments and nucleobases or their mimetics
on the other side, connected by an 1-ethylene-1,2,3-triazole
moiety. The synthetic strategy is shown in Scheme 1 and
relies on nucleophilic azidation of phosphonates 11 and 12
followed by the 1,3-dipolar cycloaddition of the azidophos-
phonates 13 and 14 with the selected alkynes.
In the key step of the synthesis of the title compounds the
1,3-dipolar cycloaddition of azidophosphonates 13 and 14
with the terminal alkynes 15a–l and 16a–c was successfully
applied.
Three groups of compounds were selected as terminal
alkynes and our collection included propargylated natural
nucleobases (N9-propargyladenine 15a, N1-propargylthymine
15b, N1-propargyluracil 15c and N4-acetyl-N1-propargylcytosine
15d), their close structural analogs (N1-propargyl-6-azauracil
15e, N1-propargyltheobromine 15f, N7-propargyltheophylline
15g, 8-chloro-N7-propargyltheophylline 15h), some heterocyclic
compounds (5,6-dimethyl-N1-propargylbenzimidazole 15i, N-
propargylmorpholine 15j, 3-acetyl-N-propargylindole 15k, N-
propargyl-2-pyridon 15l), or even fluorinated benzene deriva-
tives (1-ethynyl-2-fluorobenzene 16a, 1-ethynyl-3-fluorobenzene
16b, 1-ethynyl-2,4-difluorobenzene 16c). Under standard con-
ditions the cycloadditions were accomplished in 48–72 h.
The reaction time was shortened to 10 min when the reaction
mixtures were irradiated in a microwave oven [58] at
45–508C. In both circumstances the required cycloadducts
were obtained in comparable (60–90%) yields (Scheme 3).
Structure and purity of all diethyl ester phosphonates were
established by 1H, 31P, and 13C NMR and IR techniques as well
as by elemental analysis.
Results and discussion
Chemistry
The starting diethyl 2-chloroethoxymethylphosphonate 11
[54] and diethyl 2-chloroethoxyethylphosphonate 12 [55]
were prepared according to the literature procedures in
83% and 80% yields, respectively. The conversion of phos-
phonate 11 to azidophosphonate was carried out by reaction
with sodium azide in the presence of tetrabutylammonium
bromide in toluene at 908C to give diethyl 2-azidoethoxy-
methylphosphonate 13 [56, 57] in 73% yield. Diethyl 2-azido-
ethoxyethylphosphonate 14 was prepared according to a
similar procedure from diethyl 2-chloroethoxyethylphos-
phonate 12 in 78% yield (Scheme 2).
Finally, using bromotrimethylsilane [59] selected diethyl
phosphonates 17d, 17f, 17h, 17i, and 19a–c were transformed
into the respective phosphonic acids 22d, 22f, 22h, 22i, and
21a–c in good yields (Scheme 4). However, in the case of
the N4-acetylcytosine derivatives 17d dealkylation of the di-
ethoxyphosphoryl function was accompanied by the removal
of the acetyl group from the protected cytosine.
a
(EtO)2(O)P
N3
(EtO)2(O)P
Cl
O
n
O
n
n = 0 11
n = 1 12
n = 0 13
n = 1 14
Scheme 2. Reagents and conditions: (a) NaN3, Bu4NBr, toluene,
908C, n ¼ 0: 4 h, n ¼ 1: 24 h.
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

4
I. E. Głowacka et al.
Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
B
N
B
a
(EtO)2(O)P
N3
(EtO)2(O)P
N
N
HC C
O
n
O
n
_
_
n = 0 13
n = 1 14
15a l
n = 0 17a l
_
n = 1 18a l
CH3
NHAc
N
O
CH3
N
NH2
N
N
O
N
b
O
N
e
O
O
H3C
N
N
N
N
N
NH
O
NH
O
NH
O
N
N
N
B =
N
N
N
O
N
CH3
O
O
g
CH3
a
c
d
f
CH3
Ac
O
CH3
CH3
O
N
N
N
N
Cl
N
N
N
N
N
O
CH3
O
l
h
j
k
i
R
N
N
a
(EtO)2(O)P
N
(EtO)2(O)P
N3
HC C R
O
n
O
n
_
_
n = 0 19a c
16a c
n = 0 13
n = 1 14
_
n = 1 20a c
F
F
F
,
b
,
c
a
F
R =
Scheme 3. Reagents and conditions: (a) CuSO4 ꢁ 5H2O (0.05 equiv.), sodium ascorbate (0.1 equiv.), H2O–t-BuOH (1:1), MW, 10 min
or r.t., 48–72 h.
Antiviral activity evaluation
lung (HEL) cell cultures: herpes simplex virus-1 (KOS), herpes
simplex virus-2 (G), herpes simplex virus-1 (TKꢀ KOS ACVr),
vaccinia virus, and vesicular stomatitis virus; (b) Vero cell
cultures: para-influenza-3 virus, reovirus-1, Sindbis virus,
All synthesized compounds were evaluated for their antiviral
activities against a wide variety of DNA and RNA viruses,
using the following cell-based assays: (a) human embryonic
R
R
m
m
N
N
N
a
N
N
O
N
(HO)2(O)P
(EtO)2(O)P
O
_
_
m = 0 19a c
m = 0 21a c
m = 1 17d, f, h, i
m = 1 22d, f, h, i
CH3
NHAc
N
O
CH3
N
O
CH3
CH3
F
F
F
N
N
N
N
N
N
Cl
,
b
,
,
f
,
CH3
,
N
h
i
a
c
d
,
R =
F
N
N
O
N
O
O
CH3
Scheme 4. Reagents and conditions: (a) TMSBr, CH2Cl2, r.t., 24–48 h.
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
Synthesis and Biological Evaluation of 1,2,3-Triazolonucleotides
5
Coxsackie virus B4, Punta Toro virus; (c) HeLa cell cultures: Conclusion
vesicular stomatitis virus, Coxsackie virus B4 and respiratory
syncytial virus; (d) Crandell-Rees Feline Kidney (CRFK) cell
cultures: feline corona virus (FIPV) and feline herpes virus
(FHV); (e) Madin Darby Canine Kidney (MDCK) cell cultures:
influenza A virus H1N1 subtype (A/PR/8), influenza A virus
H3N2 subtype (A/HK/7/87) and influenza B virus (B/HK/5/72).
Ganciclovir, cidofovir, acyclovir, brivudin, (S)-9-(2,3-dihydrox-
ypropyl)adenine [(S)-DHPA], ribavirin, oseltamivir carboxy-
late, amantadine, rimantadine, and dextran sulfate
(molecular weight 5000, DS-5000) were used as the reference
The phosphonate analogs of nucleosides 17–18a–l and 19–
20a–c were synthesized from azidophosphonates 13 and 14
by the copper(I)-catalyzed Huisgen 1,3-dipolar cycloaddition
preferentially under microwave irradiation with selected
alkynes 16a–l and 17a–c, including N-propargyl nucleobases
(N9-propargyladenine 16a, N1-propargylthymine 16b, N1-prop-
argyluracil 16c, and N4-acetyl-N1-propargylcytosine 16d) and
several mimetics (N1-propargyl-6-azauracil 16e, N1-propar-
gyltheobromine 16f, N7-propargyltheophylline 16g, 8-
chloro-N7-propargyltheophylline 16h, 5,6-dimethyl-N1-prop-
argylbenzimidazole 16i, N-propargylmorpholine 16j, 3-ace-
tyl-N-propargylindole 16k, N-propargyl-2-pyridon 16l and 1-
ethynyl-2-fluorobenzene 17a, 1-ethynyl-3-fluorobenzene 17b,
1-ethynyl-2,4-difluorobenzene 17c).
compounds. The antiviral activity was expressed as the EC50
:
the compound concentration required to reduce virus-
induced cytopathogenicity by 50% (other viruses). Among
the 1,2,3-triazole derivatives, none of the compounds possess
inhibitory activity toward any of the viruses tested at sub-
toxic concentrations.
Under standard conditions (TMSBr, ethanol) some O,O-
diethylphosphonates 17d, 17f, 17h, 17i, and 19a–c were trans-
formed into their respective phosphonic acid.
Evaluation of cytotoxicity and cytostatic activity
The cytotoxicity of the tested compounds toward the unin-
fected host cells was defined as the minimum cytotoxic
All synthesized compounds were tested for their antiviral
activities against DNA and RNA viruses as well as cytostatic
activity or cytotoxicity. None of the compounds showed
inhibitory activity against virus-induced cytopathicity.
Compound 17k exhibited a modest inhibitory effect on the
proliferation of human T-lymphocyte cells at a concentration
lower than 100 mM (IC50 ¼ 64 ꢂ 13 mM for CEM).
concentration (MCC) that causes
a
microscopically
detectable alteration of normal cell morphology. The 50%
cytotoxic concentration (CC50), causing a 50% decrease in
cell viability was determined using a colorimetric 3-(4,5-
dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfo-
phenyl)-2H-tetrazolium (MTS) assay system. None of the tested
compounds affected the cell morphology of HEL, Vero, HeLa,
CRFK, and MDCK cells (MCC or CC50) at compound concen-
trations up to 100 mM.
None of the tested compounds appeared cytotoxic towards
Vero, HEL, HeLa, CRFK, and MDCK monolayer cell cultures at
concentrations up to 100–250 mM.
The cytostatic activity of the tested compounds was defined
as the 50% cytostatic inhibitory concentration (IC50). Several
compounds showed very moderately inhibitory against the
proliferation of tumor cell lines at a concentration below
Experimental
General
1H NMR spectra were taken in CDCl3, CD3OD, or D2O on the
following spectrometers: Varian Mercury-300 and Bruker Avance
III (600 MHz) with TMS as an internal standard; chemical shifts d
in ppm with respect to TMS; coupling constants J in Hz. 13C NMR
spectra were recorded for CDCl3, CD3OD, or D2O solutions on a
Varian Mercury-300 and Bruker Avance III (600 MHz) spec-
trometer at 75.5 and 150.5 MHz, respectively. 31P NMR spectra
were taken in CDCl3, CD3OD, or D2O on Varian Mercury-300 at
121.5 MHz. IR spectral data were measured on an Infinity MI-60
FT-IR spectrometer. Melting points were determined on a Boetius
apparatus and are uncorrected. Elemental analyses were
performed by the Microanalytical Laboratory of this Faculty
on a Perkin Elmer PE 2400 CHNS analyzer. The following adsorb-
ents were used: column chromatography, Merck silica gel 60
(70–230 mesh); analytical TLC, Merck TLC plastic sheets silica gel
60 F254. TLC plates were developed in chloroform–methanol
solvent systems. Visualization of spots was effected with iodine
vapours. All solvents were purified by methods described in the
literature.
250 mM (Table 1). In particular, 17k was most inhibitory (IC50
64 ꢂ 13 mM).
:
Table 1. Inhibitory effect of several 1,2,3-triazoles against the
proliferation of murine leukemia (L1210), human T-lymphocyte
(CEM), and human cervix cells (HeLa).
Compound
IC50 (mM)
L1210
CEM
HeLa
19b
17k
20c
20b
20a
18k
165 ꢂ 34
205 ꢂ 52
ꢃ250
ꢃ250
64 ꢂ 13
>250
205 ꢂ 25
148 ꢂ 7
175 ꢂ 37
201 ꢂ 25
203 ꢂ 30
121 ꢂ 15
168 ꢂ 15
180 ꢂ 13
147 ꢂ 3
>250
170 ꢂ 59
ꢃ50
All microwave irradiation experiments were carried out in
microwave reactor Plazmartonika RM 800. The reaction was
carried out in a 50 mL glass vial.
IC50 (mM), 50% inhibitory concentration.
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

6
I. E. Głowacka et al.
Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
Propargylated nucleobases and several mimetics of bases were
silica gel column with chloroform–methanol mixtures (20:1 or
10:1 v/v) to give the desired 1,2,3-triazoles.
prepared according to the literature. Uracil, thymine, adenine,
N4-acetylcytosine, 6-azauracil, theobromine, theophylline, 8-
chlorotheophylline, 2-pyridone, morpholine, and 3-acetylindole
were treated with propargyl bromide in the presence of potass-
ium carbonate in DMF to give propargylated derivatives 15a–l,
respectively [10, 13, 60–67]. In the case of synthesis of propargyl-
5,6-dimethylbenzimidazole the 5,6-dimethylbenzimidazole was
treated with propargyl bromide in the presence of tetrabutylam-
monium iodine in 50% aq. NaOH [65].
Method B
To a solution of azidophosphonate (1.00 mmol) in t-BuOH (1 mL)
and H2O (1 mL) were added CuSO4 ꢁ 5H2O (0.05 mmol), sodium
ascorbate (0.10 mmol) and alkynes (1.00 mmol). The suspension
was microwave irradiated in the microwave reactor (Plazmatronika
RM 800, 800 W) at 45–508C for 10 min. After cooling the solvent
was removed by vacuum evaporation. The residue was suspended
in chloroform (5 mL) and filtered through a layer of Celite. The
solution was concentrated in vacuo and the crude product was
purified on a silica gel column with chloroform–methanol mix-
tures (50:1, 20:1 or 10:1 v/v) to give the desired 1,2,3-triazoles.
1-Ethynyl-2,4-difluorobenzene, 1-ethynyl-2-fluorobenzene, and
1-ethynyl-3-fluorobenzene were commercially available.
Synthesis of 2-azidoethoxymethylphosphonate 13
A suspension of 2-chloroethoxymethylphosphonate 11 (8.743 g,
37.910 mmol),
tetrabutylammonium
bromide
(1.833 g,
Diethyl {2-[4-((6-amino-9H-purin-9-yl)methyl)-1H-1,2,3-
triazol-1-yl]ethoxy}methylphosphonate 17a
5.686 mmol) as a phase transfer catalyst and sodium azide
(4.683 g, 72.030 mmol) in (50 mL) of toluene was heated to
908C for 4 h. After cooling, the suspension was filtered through
a layer of Celite. The solution was evaporated in vacuo and the
crude product was purified on a silica gel column with chloro-
form–methanol mixtures (100:1 v/v) to give pure diethyl 2-azi-
doethoxymethylphosphonate 13 (6.583 g, 73%) as colorless oil; IR
(film, cmꢀ1) nmax: 2968, 2910, 2878, 2158, 1225, 1025; 1H NMR
(300 MHz, CDCl3) d: 1.35 (t, J ¼ 7.2 Hz, 6H, 2 ꢄ POCH2CH3), 3.41
(t, J ¼ 5.2 Hz, 2H, OCH2CH2), 3.77 (t, J ¼ 5.2 Hz, 2H, PCH2), 3.85
(d, J ¼ 8.1 Hz, 2H, OCH2CH2), 4.14–4.24 (m, 4H, 2 ꢄ POCH2CH3);
13C NMR (75.5 MHz, CDCl3) d: 16.6 (d, J ¼ 5.3 Hz, POCC), 50.8, 62.8
(d, J ¼ 6.6 Hz, 2 ꢄ POC), 65.2 (d, J ¼ 166.0 Hz, PC), 72.2 (d,
J ¼ 10.8 Hz, OCH2CH2); 31P NMR (121.5 MHz, CDCl3) d ¼ 21.64.
According to the general procedure (method B) from the azide 13
(0.300 g, 1.26 mmol) and N9-propargyladenine 15a (0.219 g,
1.26 mmol) a phosphonate 17a (0.443 g, 86%) was obtained after
chromatography on a silica gel column with chloroform–
methanol (10:1 v/v) as a white powder, m.p.: 94–968C; IR
(KBr, cmꢀ1) nmax: 3468, 3300, 3121, 2987, 2918, 1661, 1601,
1471, 1229, 1050, 1025; 1H NMR (300 MHz, CDCl3) d: 1.27
(t, J ¼ 7.0 Hz, 6H, 2 ꢄ POCH2CH3), 3.74 (d, J ¼ 8.1 Hz, 2H,
PCH2), 3.96 (t, J ¼ 5.0 Hz, 2H, OCH2CH2), 4.03–4.17 (m, 4H,
2 ꢄ POCH2CH3), 4.54 (d, J ¼ 5.0 Hz, 2H, OCH2CH2), 5.48 (s, 2H,
0
CH2), 6.02 (brs, 2H, NH2), 7.84 (s, 1H, HC5 ) 8.00 (s, 1H, NCHN),
8.36 (s, 1H, NCHN); 13C NMR (75.5 MHz, CDCl3) d: 16.6
(d, J ¼ 5.4 Hz, POCC), 38.6, 50.4, 62.7 (d, J ¼ 6.6 Hz, 2 ꢄ POC), 65.2
(d, J ¼ 166.3 Hz, PC), 71.2 (d, J ¼ 10.1 Hz, OCH2CH2), 119.4, 124.1
(s, HC ¼ C), 140.4 (s, HC ¼ C), 142.3, 149.7, 152.9, 155.7; 31P NMR
(121.5 MHz, CDCl3) d: 21.41; Anal. calcd. for C15H23N8O4P: C, 43.90;
H, 5.65; N, 27.31. Found: C, 44.14; H, 5.81; N, 27.18.
Synthesis of 2-azidoethoxyethylphosphonate 14
A suspension of 2-chloroethoxyethylphosphonate 12 (3.004 g,
12.279 mmol), tetrabutylammonium bromide (0.594 g, 1.840 mmol)
as
a phase transfer catalyst and sodium azide (1.517 g,
23.329 mmol) in (20 mL) of toluene was heated to 908C for
24 h. After cooling, the suspension was filtered through a layer
of Celite. The solution was evaporated in vacuo and the crude
product was purified on a silica gel column with chloroform–
methanol mixtures (100:1 v/v) to give pure diethyl 2-azidoethoxy-
Diethyl {2-[4-((5-methyl-2,4-dioxopyrimidin-1-yl)methyl)-
1H-1,2,3-triazol-1-yl]ethoxy}methylphosphonate 17b
According to the general procedure (method B) from the azide 13
(0.200 g, 0.840 mmol) and N1-propargylthymine 15b (0.138 g,
0.840 mmol) a phosphonate 17b (0.366 g, 90%) was obtained
after chromatography on a silica gel column with chloro-
form–methanol (10:1 v/v) as a white, amorphous solid, m.p.:
71–738C; IR (KBr, cmꢀ1) nmax: 3448, 3147, 3037, 2987, 2926,
2831, 1686, 1242, 1027, 970; 1H NMR (300 MHz, CDCl3) d: 1.32
(t, J ¼ 7.0 Hz, 6H, 2 ꢄ POCH2CH3), 1.90 (d, J ¼ 1.2 Hz, 3H,
ethylphosphonate 14 (2.395 g, 78%) as colourless oil; IR (film, cmꢀ1
)
nmax: 2891, 2870, 2813, 2169, 1245, 1030; 1H NMR (600 MHz,
CDCl3) d: 1.35 (t, J ¼ 7.0 Hz, 6H, 2 ꢄ POCH2CH3), 2.14
(dt, J ¼ 18.7 Hz, J ¼ 7.6 Hz, 2H, PCH2), 3.40 (t, J ¼ 5.2 Hz, 2H,
OCH2CH2), 3.65 (t, J ¼ 5.2 Hz, 2H, OCH2CH2), 3.76 (dt, J ¼ 11.3 Hz,
J ¼ 7.6 Hz, 2H, PCH2CH2), 4.07–4.17 (m, 4H, 2 ꢄ POCH2CH3);
13C NMR (75.5 MHz, CDCl3) d: 16.4 (d, J ¼ 6.0 Hz, POCC), 27.0
(d, J ¼ 139.4 Hz, PC), 50.6, 61.7 (d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.1,
69.5; 31P NMR (121.5 MHz, CDCl3) d ¼ 28.11.
–
HC CCH ), 3.79 (d, J ¼ 7.8 Hz, 2H, PCH ), 4.00 (t, J ¼ 5.1 Hz, 2H,
–
3
2
OCH2CH2), 4.06–4.18 (m, 4H, 2 ꢄ POCH2CH3), 4.57 (t, J ¼ 5.1 Hz,
–
–
2H, OCH CH ), 4.96 (s, 2H, CH ), 7.36 (d, J ¼ 1.2 Hz, 1H, HC CCH ),
2
2
2
3
7.86 (s, 1H, HC5 ), 8.91 (brs, 1H, NH); 13C NMR (75.5 MHz, CDCl3)
d: 12.5, 16.6 (d, J ¼ 6.3 Hz, POCC), 42.8, 50.2, 62.6 (d, J ¼ 6.3 Hz,
0
–
2 ꢄ POC), 65.3 (d, J ¼ 130.8 Hz, PC), 111.1, 124.7 (s, HC C), 140.2,
–
General procedure for the preparation of 1,2,3-triazoles
142.0 (s, HC C), 151.0 (s, C O), 164.5 (s, C O); 31P NMR (121.5 MHz,
–
–
–
–
–
–
CDCl3) d: 21.42; Anal. calcd. for C15H24N5O6P: C, 44.89; H, 6.03;
N, 17.45. Found: C, 44.66; H, 5.83; N, 17.37.
Method A
To a solution of azidophosphonate (1.00 mmol) in t-BuOH (0.5 mL)
and H2O (1 mL) were added CuSO4 ꢁ 5H2O (0.05 mmol), sodium
ascorbate (0.10 mmol) followed by alkynes (1.00 mmol). The sus-
pension was vigorously stirred at room temperature for 48–72 h.
After removal of solvents the residue was suspended in chloroform
(5 mL) and filtered through a layer of Celite. The solution was
concentrated in vacuo and the crude product was purified on a
Diethyl {2-[4-((2,4-dioxopyrimidin-1-yl)methyl)-1H-
1,2,3-triazol-1-yl]ethoxy}methylphosphonate 17c
According to the general procedure (method B) from the azide 13
(0.200 g, 0.840 mmol) and N1-propargyluracil 15c (0.138 g,
0.840 mmol) a phosphonate 17c (0.366 g, 78%) was obtained after
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
Synthesis and Biological Evaluation of 1,2,3-Triazolonucleotides
7
chromatography on a silica gel column with chloroform–
0.675 mmol) a phosphonate 17f (0.233 g, 76%) was obtained after
chromatography on a silica gel column with chloroform–
methanol (50:1, 20:1 v/v) as a colorless oil.
methanol (10:1 v/v) as a colorless oil. IR (film, cmꢀ1) nmax
:
3150, 2987, 2824, 1680, 1459, 1238, 1048, 811, 785; 1H NMR
(300 MHz, CDCl3) d: 1.31 (t, J ¼ 7.1 Hz, 6H, 2 ꢄ POCH2CH3), 3.79
(d, J ¼ 8.1 Hz, 2H, PCH2), 4.00 (t, J ¼ 5.0 Hz, 2H, OCH2CH2), 4.09–
4.17 (m, 4H, 2 ꢄ POCH2CH3), 4.57 (t, J ¼ 5.0 Hz, 2H, OCH2CH2),
According to the general procedure (method B) from the azide
13 (0.160 g, 0.675 mmol) and N1-propargyltheobromine 15f
(0.147 g, 0.675 mmol) a phosphonate 17f (0.251 g, 82%) was
obtained after chromatography on a silica gel column with
chloroform–methanol (50:1, 20:1 v/v) as a colorless oil; IR
(film, cmꢀ1) nmax: 3113, 2983, 2905, 1707, 1663, 1236, 1026,
974; 1H NMR (600 MHz, CDCl3) d: 1.34 (t, J ¼ 7.1 Hz, 6H,
2 ꢄ POCH2CH3), 3.59 (s, 3H, N–CH3), 3.79 (d, J ¼ 8.2 Hz, 2H,
PCH2), 3.99 (t, J ¼ 5.3 Hz, 2H, OCH2CH2), 4.01 (s, 3H, N–CH3),
4.02–4.12 (m, 4H, 2 ꢄ POCH2CH3), 4.53 (t, J ¼ 5.3 Hz, 2H,
OCH2CH2), 5.34 (s, 2H, CH2), 7.52 (s, 1H), 7.76 (s, 1H, HC5’);
13C NMR (151 MHz, CDCl3) d: 16.4 (d, J ¼ 5.5 Hz, POCC),
29.6, 33.5, 35.9, 49.9, 62.4 (d, J ¼ 6.5 Hz, 2 ꢄ POC), 65.2
(d, J ¼ 166.5 Hz, PC), 71.2 (d, J ¼ 10.2 Hz, OCH2CH2), 107.5,
–
–
4.99 (s, 2H, CH2), 5.70 (d, J ¼ 7.9 Hz, 1H, HC CH), 7.53
–
–
0
(d, J ¼ 7.9 Hz, 1H, HC CH), 7.89 (s, 1H, HC ), 9.20 (brs, 1H,
5
NH); 13C NMR (75.5 MHz, CDCl3) d: 16.7 (d, J ¼ 5.7 Hz, POCC),
43.1, 50.3, 62.8 (d, J ¼ 6.6 Hz, 2 ꢄ POC), 65.3 (d, J ¼ 166.3 Hz,
–
–
PC), 71.2 (d, J ¼ 10.0 Hz, OCH CH ), 102.7, 124.9 (s, HC C), 141.8
2
2
(s, HC C), 144.5, 151.0 (s, C O), 164.0 (s, C O); 31P NMR
–
–
–
–
–
–
(121.5 MHz, CDCl3) d: 21.48; Anal. calcd. for C14H22N5O6P: C,
43.41; H, 5.73; N, 18.08. Found: C, 43.66; H, 5.88; N, 17.97.
Diethyl {2-[4-((4-acetylamino-2-oxopyrimidin-1-yl)methyl)-
1H-1,2,3-triazol-1-yl]ethoxy}methylphosphonate 17d
According to the general procedure (method B) from the azide 13
(0.189 g, 0.797 mmol) and N4-acetyl-N1-propargylcytosine 15d
(0.152 g, 0.797 mmol) a phosphonate 17d (0.241 g, 71%) was
obtained after chromatography on a silica gel column with
chloroform–methanol (50:1, 20:1, 10:1 v/v) as a white, amor-
phous solid, m.p.: 124–1268C; IR (KBr, cmꢀ1) nmax: 3448, 3223,
3137, 2980, 2903, 1711, 1650, 1561, 1235, 1051, 1017, 795;
1H NMR (300 MHz, CDCl3) d: 1.31 (t, J ¼ 6.7 Hz, 6H,
2 ꢄ POCH2CH3), 2.24 (s, 3H, C(O)CH3), 3.79 (d, J ¼ 8.1 Hz, 2H,
PCH2), 4.00 (t, J ¼ 5.2 Hz, 2H, OCH2CH2), 4.07–4.14 (m, 4H,
2 ꢄ POCH2CH3), 4.55 (t, J ¼ 5.2 Hz, 2H, OCH2CH2), 5.14 (s, 2H,
–
–
–
124.1 (s, HC C), 141.7 (s, HC C), 143.5, 148.8, 151.2, 154.7;
–
31P NMR (243 MHz, CDCl3) d: 20.32; Anal. calcd. for
C17H26N7O6P: C, 44.84; H, 5.75; N, 21.53. Found: C, 45.02; H,
5.88; N, 21.49.
Diethyl {2-[4-((1,3-dimethyl-2,6-dioxopurin-7-yl)methyl)-
1H-1,2,3-triazol-1-yl]ethoxy}methylphosphonate 17g
According to the general procedure (method B) from the azide 13
(0.120 g, 0.506 mmol) and N7-propargyltheophylline 15g
(0.110 g, 0.506 mmol) a phosphonate 17g (0.189 g, 82%) was
obtained after chromatography on a silica gel column with
chloroform–methanol (50:1, 20:1 v/v) as a white solid, m.p.:
124–1258C; IR (KBr, cmꢀ1) nmax: 3110, 2964, 1705, 1659, 1257,
1050, 760; 1H NMR (600 MHz, CDCl3) d: 1.33 (t, J ¼ 7.1 Hz, 6H,
2 ꢄ POCH2CH3), 3.43 (s, 3H, N–CH3), 3.59 (s, 3H, N–CH3), 3.78
(d, J ¼ 8.0 Hz, 2H, PCH2), 4.01 (t, J ¼ 5.0 Hz, 2H, OCH2CH2), 4.09–
4.14 (m, 4H, 2 ꢄ POCH2CH3), 4.56 (t, J ¼ 5.0 Hz, 2H, OCH2CH2),
–
0
5
CH ), 7.38 (d, J ¼ 7.8 Hz, 1H, HC CH), 7.94 (s, 1H, HC ), 7.95
–
2
(d, J ¼ 7.3 Hz, 1H, HC CH), 9.31 (brs, 1H, NH); 13C NMR (75.5 MHz,
–
–
CDCl3) d: 16.5 (d, J ¼ 6.0 Hz, POCC), 24.8 (s, C(O)CH3), 44.9, 50.0,
62.5 (d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.0 (d, J ¼ 165.6 Hz, PC), 71.0
–
–
–
–
(d, J ¼ 10.2 Hz, OCH CH ), 97.0, 124.9 (s, HC C), 141.6 (s, HC C),
2
2
–
148.9, 155.6, 162.9 (s, C O), 171.1 (s, C O); 31P NMR (121.5 MHz,
–
–
–
5.61 (s, 2H, CH2), 7.82 (s, 1H), 7.95 (s, 1H, HC5 ); 13C NMR (151 MHz,
0
CDCl3) d: 21.50; Anal. calcd. for C16H25N6O6P: C, 44.86; H, 5.88;
N, 19.62. Found: C, 44.68; H, 5.63; N, 19.37.
CDCl3) d: 16.4 (d, J ¼ 5.5 Hz, POCC), 27.9, 29.7, 41.4, 50.1, 62.4
(d, J ¼ 6.6 Hz, 2 ꢄ POC), 65.4 (d, J ¼ 166.6 Hz, PC), 71.0
–
–
–
(d, J ¼ 9.7 Hz, OCH CH ), 106.5, 124.5 (s, HC C), 141.3 (s, HC C),
–
2
2
142.1, 148.9, 151.6, 155.3; 31P NMR (243 MHz, CDCl3) d: 20.24;
Anal. calcd. for C17H26N7O6P: C, 44.84; H, 5.75; N, 21.53. Found: C,
44.62; H, 5.78; N, 21.60.
Diethyl {2-[4-((3,5-dioxo-1,2,4-triazin-2-yl)methyl)-1H-
1,2,3-triazol-1-yl]ethoxy}methyl phosphonate 17e
According to the general procedure (method B) from the azide 13
(0.126 g, 0.531 mmol) and N1-propargyl-6-azauracil 15e (0.080 g,
0.531 mmol) a phosphonate 17e (0.183 g, 93%) was obtained after
chromatography on a silica gel column with chloroform–
Diethyl {2-[4-((8-chloro-1,3-dimethyl-2,6-dioxopurin-7-yl)-
methyl)-1H-1,2,3-triazol-1-yl]ethoxy}methylphosphonate 17h
According to the general procedure (method B) from the azide 13
(0.160 g, 0.675 mmol) and 8-chloro-N7-propargyltheophylline
15h (0.170 g, 0.675 mmol) a phosphonate 17h (0.251 g, 76%)
was obtained after chromatography on a silica gel column
with chloroform–methanol (50:1, 20:1 v/v) as a colorless oil;
IR (film, cmꢀ1) nmax: 3140, 2986, 2887, 1707, 1664, 1237, 1029,
751; 1H NMR (600 MHz, CDCl3) d: 1.34 (t, J ¼ 7.0 Hz, 6H,
2 ꢄ POCH2CH3), 3.43 (s, 3H, N–CH3), 3.56 (s, 3H, N–CH3), 3.78
(d, J ¼ 8.1 Hz, 2H, PCH2), 4.00 (t, J ¼ 5.0 Hz, 2H, OCH2CH2), 4.10–
4.18 (m, 4H, 2 ꢄ POCH2CH3), 4.55 (t, J ¼ 5.0 Hz, 2H, OCH2CH2),
methanol (50:1, 20:1, 10:1 v/v) as a colorless oil; IR (film, cmꢀ1
)
nmax: 3220, 3109, 3020, 2918, 1734, 1678, 1430, 1226, 1023,
758; 1H NMR (300 MHz, CDCl3) d: 1.32 (t, J ¼ 7.0 Hz,
6H, 2 ꢄ POCH2CH3), 3.79 (d, J ¼ 7.3 Hz, 2H, PCH2), 3.97
(t, J ¼ 5.0 Hz, 2H, OCH2CH2), 4.08–4.18 (m, 4H, 2 ꢄ POCH2CH3),
4.53 (t, J ¼ 5.0 Hz, 2H, OCH2CH2), 5.20 (s, 2H, CH2), 7.40 (s, 1H), 7.82
(s, 1H, HC5’), 10.66 (brs, 1H, NH); 13C NMR (75.5 MHz, CDCl3) d: 16.6
(d, J ¼ 5.8 Hz, POCC), 34.7, 50.2, 62.9 (d, J ¼ 6.6 Hz, 2 ꢄ POC),
65.1 (d, J ¼ 166.6 Hz, PC), 71.3 (d, J ¼ 10.6 Hz, OCH2CH2), 124.8
(s, HC C), 134.8, 141.6 (s, HC C), 149.1 (s, C O), 155.9 (s, C O); 31P NMR
–
–
–
–
–
–
–
–
(121.5 MHz, CDCl3) d: 21.59; Anal. calcd. for C13H21N6O6P: C, 40.21;
H, 5.45; N, 21.64. Found: C, 40.48; H, 5.23; N, 21.83.
5.66 (s, 2H, CH2), 7.91 (s, 1H, HC5 ); 13C NMR (151 MHz, CDCl3)
0
d: 16.4 (d, J ¼ 5.5 Hz, POCC), 28.0, 29.8, 41.0, 50.2, 62.4
(d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.4 (d, J ¼ 166.8 Hz, PC), 71.1
Diethyl {2-[4-((3,7-dimethyl-2,6-dioxopurin-1-yl)methyl)-
–
(d, J ¼ 9.8 Hz, OCH CH ), 107.5, 124.4 (s, HC C), 139.0, 141.7
–
2
2
(s, HC C), 147.4, 151.3, 154.5; 31P NMR (243 MHz, CDCl3)
d: 20.26; Anal. calcd. for C17H25ClN7O6P: C, 41.68; H, 5.14;
N, 20.02. Found: C, 41.80; H, 5.23; N, 19.97.
–
–
1H-1,2,3-triazol-1-yl]ethoxy}methylphosphonate 17f
According to the general procedure (method A) from the azide 13
(0.160 g, 0.675 mmol) and N1-propargyltheobromine 15f (0.147 g,
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

8
I. E. Głowacka et al.
Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
PCH2), 3.95 (t, J ¼ 5.1 Hz, 2H, OCH2CH2), 3.99–4.09 (m, 4H,
2 ꢄ POCH2CH3), 4.52 (t, J ¼ 5.1 Hz, 2H, OCH2CH2), 5.47 (s, 2H,
CH2), 7.27–7.44 (m, 2H, Haromat), 7.42–7.48 (m, 1H, Haromat), 7.67
Diethyl {2-[4-((5,6-dimethyl-1H-benzo[d]imidazol-1-yl)-
methyl)-1H-1,2,3-triazol-1-yl]ethoxy}methylphosphonate 17i
According to the general procedure (method A) from the azide 13
(0.160 g, 0.675 mmol) and 5,6-dimethyl-N1-propargylbenzimid-
azole 15i (0.124 g, 0.675 mmol) a phosphonate 17i (0.177 g,
62%) was obtained after chromatography on a silica gel column
with chloroform–methanol (50:1, 20:1 v/v) as a colorless oil.
According to the general procedure (method B) from the azide
13 (0.145 g, 0.611 mmol) and N1-propargyl-5,6-dimethylbenzimi-
dazole 15i (0.113 g, 0.611 mmol) a phosphonate 17i (0.196 g,
76%) was obtained after chromatography on a silica gel column
with chloroform–methanol (50:1, 20:1 v/v) as a colorless oil;
IR (film, cmꢀ1) nmax: 3135, 3093, 2982, 2936, 1227, 1049, 972,
830; 1H NMR (300 MHz, CDCl3) d: 1.28 (t, J ¼ 7.0 Hz, 6H,
2 ꢄ POCH2CH3), 2.39 (s, 3H, CH3), 2.40 (s, 3H, CH3), 3.72
(d, J ¼ 8.1 Hz, 2H, PCH2), 3.97 (t, J ¼ 5.0 Hz, 2H, OCH2CH2),
4.02–4.10 (m, 4H, 2 ꢄ POCH2CH3), 4.53 (t, J ¼ 5.0 Hz, 2H,
OCH2CH2), 5.49 (s, 2H, CH2), 7.31, 7.59, 7.67 (s, 1H), 8.13 (s, 1H,
(s, 1H), 7.92 (s, 1H, HC5 ), 8.35–8.40 (m, 1H, Haromat); 13C NMR
0
(75.5 MHz, CDCl3) d: 16.6 (d, J ¼ 5.4 Hz, POCC), 27.8, 42.4, 50.3,
62.6 (d, J ¼ 6.3 Hz, 2 ꢄ POC), 64.6 (d, J ¼ 166.9 Hz, PC), 71.2
(d, J ¼ 10.3 Hz, OCH2CH2), 109.9, 117.5, 122.7, 122.7, 123.5,
123.6, 126.4, 134.8, 136.6, 142.8, 193.0; 31P NMR (121.5 MHz,
CDCl3) d: 20.68; Anal. calcd. for C20H27N4O5P: C, 55.29; H, 6.26;
N, 12.90. Found: C, 55.46; H, 6.33; N, 12.81.
Diethyl {2-[4-((2-oxopyridin-1-yl)methyl)-1H-1,2,3-triazol-
1-yl]ethoxy}methylphosphonate 17l
According to the general procedure (method B) from the azide 13
(0.131 g, 0.552 mmol) and N1-propargyl-2-pyridone 15l (0.074 g,
0.552 mmol) a phosphonate 17l (0.149 g, 73%) was obtained after
chromatography on a silica gel column with chloroform–
methanol (50:1, 20:1 v/v) as a greyish oil; IR (film, cmꢀ1) nmax
:
HC5 ); 13C NMR (75.5 MHz, CDCl3) d: 16.4 (d, J ¼ 5.5 Hz, POCC),
3418, 2984, 2933, 1657, 1581, 1225, 1026; 1H NMR (300 MHz,
CDCl3) d: 1.31 (t, J ¼ 7.0 Hz, 6H, 2 ꢄ POCH2CH3), 3.77
(d, J ¼ 8.0 Hz, 2H, PCH2), 3.99 (t, J ¼ 5.2 Hz, 2H, OCH2CH2),
4.00–4.17 (m, 4H, 2 ꢄ POCH2CH3), 4.52 (t, J ¼ 5.2 Hz, 2H,
OCH2CH2), 5.19 (s, 2H, CH2), 6.18 (dt, J ¼ 8.1 Hz, J ¼ 6.5 Hz,
1H, Haromat), 6.54 (ddd, J ¼ 9.2 Hz, J ¼ 1.4 Hz, J ¼ 0.8 Hz,
1H, Haromat), 7.31 (ddd, J ¼ 9.2 Hz, J ¼ 6.9 Hz, J ¼ 2.2 Hz,
1H, Haromat), 7.59 (dd, J ¼ 6.9 Hz, J ¼ 2.2 Hz, 1H, Haromat), 7.87
0
20.1, 20.5, 40.4, 50.1, 62.4 (d, J ¼ 6.5 Hz, 2 ꢄ POC), 65.2
(d, J ¼ 167.1 Hz, PC), 71.1 (d, J ¼ 9.9 Hz, OCH2CH2), 110.0,
–
–
–
120.2, 123.3 (s, HC C), 133.3, 132.1, 132.4, 142.0 (s, HC C),
–
142.3, 143.3; 31P NMR (243 MHz, CDCl3) d: 20.96; Anal. calcd.
for C19H28N5O4P: C, 54.15; H, 6.70; N, 16.62. Found: C, 54.28;
H, 6.83; N, 16.49.
(s, 1H, HC5 ); 13C NMR (75.5 MHz, CDCl3) d: 16.5 (d, J ¼ 6.0 Hz,
0
Diethyl {2-[4-(morpholinomethyl)-1H-1,2,3-triazol-1-yl]-
ethoxy}methylphosphonate 17j
According to the general procedure (method A) from the azide 13
(0.160 g, 0.675 mmol) and N1-propargylmorpholine 15j (0.084 g,
0.675 mmol) a phosphonate 17j (0.211 g, 86%) was obtained
after chromatography on a silica gel column with chloroform–
methanol (50:1, 20:1 v/v) as a colorless oil.
POCC), 44.3, 50.2, 62.6 (d, J ¼ 6.4 Hz, 2 ꢄ POC), 64.5
(d, J ¼ 166.0 Hz, PC), 71.3 (d, J ¼ 10.6 Hz, OCH2CH2), 106.5,
120.6, 124.9 (s, HC C), 137.7, 139.9, 162.2 (s, C O); 31P NMR
–
–
–
–
(121.5 MHz, CDCl3) d: 21.37; Anal. calcd. for C15H23N4O5P: C,
48.65; H, 6.26; N, 15.13. Found: C, 48.36; H, 6.14; N, 15.31.
Diethyl {2-[4-(2-fluorophenyl)-1H-1,2,3-triazol-1-yl]-
ethoxy}methylphosphonate 19a
According to the general procedure (method A) from the azide 13
(0.150 g, 0.630 mmol) and ethynyl-2-fluorobenzene 16a (0.076 g,
0.630 mmol) a phosphonate 19a (0.153 g, 68%) was obtained
after chromatography on a silica gel column with chloroform–
methanol (100:1 v/v) as a colorless oil.
According to the general procedure (method B) from the azide
13 (0.150 g, 0.630 mmol) and ethynyl-2-fluorobenzene 16a
(0.076 g, 0.630 mmol) phosphonate 19a (0.167 g, 74%) was
obtained after chromatography on a silica gel column with
According to the general procedure (method B) from the
azide 13 (0.155 g, 0.654 mmol) and N1-propargylmorpholine
15j (0.082 g, 0.654 mmol) a phosphonate 17j (0.204 g, 86%)
was obtained after chromatography on a silica gel column
with chloroform–methanol (50:1, 20:1 v/v) as a colorless oil;
IR (film, cmꢀ1) nmax: 2913, 2857, 1238, 1116, 1026; 1H NMR
(600 MHz, CDCl3) d: 1.35 (t, J ¼ 7.0 Hz, 6H, 2 ꢄ POCH2CH3),
2.54 (brt, J ¼ 4.6 Hz, 4H, CH2OCH2), 3.69 (s, 2H, CH2), 3.73
(t, J ¼ 4.6 Hz, 4H, CH2NCH2), 3.80 (d, J ¼ 8.3 Hz, 2H, PCH2), 4.01
(t, J ¼ 4.9 Hz, 2H, OCH2CH2), 4.10–4.19 (m, 4H, 2 ꢄ POCH2CH3),
4.58 (t, J ¼ 4.9 Hz, 2H, OCH2CH2), 7.68 (s, 1H, HC5 ); 13C NMR
0
chloroform–methanol (100:1 v/v) as
a colorless oil; IR
(151 MHz, CDCl3) d: 16.4 (d, J ¼ 5.6 Hz, POCC), 49.9, 53.3, 53.6,
62.5 (d, J ¼ 6.5 Hz, 2 ꢄ POC), 65.3 (d, J ¼ 166.2 Hz, PC), 66.8, 71.4
(film, cmꢀ1) nmax: 3148, 2984, 2907, 1487, 1234, 1028, 973,
818, 764; 1H NMR (600 MHz, CDCl3) d: 1.30 (t, J ¼ 7.0 Hz, 6H,
2 ꢄ POCH2CH3), 3.82 (d, J ¼ 8.4 Hz, 2H, PCH2), 4.06 (t, J ¼ 5.2 Hz,
2H, OCH2CH2), 4.10–4.17 (m, 4H, 2 ꢄ POCH2CH3), 4.67 (t, J ¼ 5.2 Hz,
2H, OCH2CH2), 7.16 (ddd, J ¼ 11.2 Hz, J ¼ 8.2 Hz, J ¼ 1.2 Hz,
1H, Haromat), 7.27 (dt, J ¼ 7.8 Hz, J ¼ 1.2 Hz, 1H, Haromat), 7.31–
(d, J ¼ 10.9 Hz, OCH CH ), 123.9 (s, HC C), 143.9 (s, HC C); 31P NMR
–
–
–
–
2
2
(243 MHz, CDCl3) d: 20.33; Anal. calcd. for C14H27N4O5P: C, 46.40;
H, 7.51; N, 15.46. Found: C, 46.58; H, 7.73; N, 15.69.
Diethyl {2-[4-((3-acetyl-1H-indol-1-yl)methyl)-1H-1,2,3-
triazol-1-yl]ethoxy}methylphosphonate 17k
0
7.36 (m, 1H, Haromat), 8.01 (d, J ¼ 3.8 Hz, 1H, HC5 ), 8.32
(dt, J ¼ 7.8 Hz, J ¼ 1.9 Hz, 1H, Haromat); 13C NMR (151 MHz,
CDCl3) d: 16.3 (d, J ¼ 6.0 Hz, POCC), 50.4 (s, OCH2CH2),
62.5 (d, J ¼ 6.6 Hz, 2 ꢄ POC), 64.6 (d, J ¼ 166.1 Hz, PC), 71.4
(d, J ¼ 10.9 Hz, OCH2CH2), 115.6 (d, J ¼ 21.3Hz), 118.6
(d, J ¼ 13.0 Hz), 123.9 (d, J ¼ 12.4 Hz), 124.6 (d, J ¼ 3.4 Hz),
127.6 (d, J ¼ 3.4 Hz), 129.3 (d, J ¼ 8.1 Hz), 141.2 (d, J ¼ 2.4 Hz),
159.1 (d, J ¼ 247.8 Hz); 31P NMR (243 MHz, CDCl3) d: 20.17;
Anal. calcd. for C15H21FN3O4P: C, 50.42; H, 5.92; N, 11.76.
Found: C, 50.56; H, 5.78; N, 11.66.
According to the general procedure (method B) from the azide 13
(0.112 g, 0.472 mmol) and 3-acetyl-N1-propargylindole 15k
(0.093 g, 0.472 mmol) a phosphonate 17k (0.173 g, 84%) was
obtained after chromatography on a silica gel column with
chloroform–methanol (50:1, 20:1 v/v) as a colorless oil; IR
(film, cmꢀ1) nmax: 3466, 3107, 2984, 2932, 1643, 1528, 1237,
1027, 752; 1H NMR (300 MHz, CDCl3) d: 1.25 (t, J ¼ 6.7 Hz, 6H,
2 ꢄ POCH2CH3), 2.53 (s, 3H, C(O)CH3), 3.72 (d, J ¼ 8.3 Hz, 2H,
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
Synthesis and Biological Evaluation of 1,2,3-Triazolonucleotides
9
chromatography on a silica gel column with chloroform–
methanol (20:1, 10:1 v/v) as a yellow oil; IR (film, cmꢀ1) nmax
Diethyl {2-[4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl]-
ethoxy}methylphosphonate 19b
According to the general procedure (method A) from the azide 13
(0.150 g, 0.630 mmol) and ethynyl-3-fluorobenzene 16b (0.076 g,
0.630 mmol) a phosphonate 19b (0.160 g, 71%) was obtained
after chromatography on a silica gel column with chloro-
form–methanol (100:1 v/v) as a colorless oil.
:
3326, 3170, 2988, 2907, 1644, 1599, 1477, 1230, 1052, 1029,
756; 1H NMR (300 MHz, CDCl3) d: 1.29 (t, J ¼ 7.0 Hz, 6H,
2 ꢄ POCH2CH3), 2.02 (dt, J ¼ 18.6 Hz, J ¼ 7.3 Hz, 2H, PCH2), 3.67
(dt, J ¼ 13.0 Hz, J ¼ 7.3 Hz, 2H, PCH2CH2), 3.79 (t, J ¼ 5.1 Hz, 2H,
OCH2CH2), 4.01–4.11 (m, 4H, 2 ꢄ POCH2CH3), 4.51 (t, J ¼ 5.1 Hz,
2H, OCH2CH2), 5.48 (s, 2H, CH2), 5.89 (brs, 2H, NH2), 7.89 (s, 1H,
According to the general procedure (method B) from the azide 13
(0.150 g, 0.630 mmol) and ethynyl-3-fluorobenzene 16b (0.076 g,
0.630 mmol) a phosphonate 19b (0.172 g, 76%) was obtained after
chromatography on a silica gel column with chloroform–methanol
(100:1 v/v) as a colorless oil; IR (film, cmꢀ1) nmax: 3123, 2970, 2943,
NCHN), 8.02 (s, 1H, HC5 ), 8.37 (s, 1H, NCHN); 13C NMR (75.5 MHz,
0
CDCl3) d: 16.5 (d, J ¼ 6.0 Hz, POCC), 27.0 (d, J ¼ 140.0 Hz, PC), 38.6,
–
50.3, 61.7 (d, J ¼ 6.6 Hz, 2 ꢄ POC), 65.2, 68.8, 119.2, 124.1 (s, HC C),
–
140.3 (s, HC C), 142.1, 149.5, 152.8, 155.8; 31P NMR (121.5 MHz,
–
–
CDCl3) d: 28.31; Anal. calcd. for C16H25N8O4P: C, 45.28; H, 5.94;
N, 26.40. Found: C, 45.52; H, 6.08; N, 26.32.
1
2813, 1482, 1227, 1050, 962, 823, 771; H NMR (600 MHz, CDCl3)
d: 1.30 (t, J ¼ 7.0 Hz, 6H, 2 ꢄ POCH2CH3), 3.82 (d, J ¼ 8.5 Hz, 2H,
PCH2), 4.05 (t, J ¼ 5.0 Hz, 2H, OCH2CH2), 4.10–4.15 (m, 4H,
2 ꢄ POCH2CH3), 4.61 (t, J ¼ 5.0 Hz, 2H, OCH2CH2), 7.00–7.06
(m, 1H, Haromat), 7.39 (dt, J ¼ 8.4 Hz, J ¼ 6.0 Hz, 1H, Haromat),
Diethyl {2-[4-((5-methyl-2,4-dioxopyrimidin-1-yl)methyl)-
1H-1,2,3-triazol-1-yl]ethoxy}methylphosphonate 18b
According to the general procedure (method B) from the azide 14
(0.100 g, 0.398 mmol) and N1-propargylthymine 15b (0.065 g,
0.398 mmol) a phosphonate 18b (0.144 g, 87%) was obtained
after chromatography on a silica gel column with chloro-
form–methanol (20:1, 10:1 v/v) as a white solid, m.p.: 65–678C;
IR (KBr, cmꢀ1) nmax: 3136, 2990, 2865, 2826, 1689, 1464, 1244,
1056, 754; 1H NMR (300 MHz, CDCl3) d: 1.32 (t, J ¼ 7.0 Hz, 6H,
7.61–7.67 (m, 2H, Haromat), 8.18 (s, 1H, HC5 ); 13C NMR (151 MHz,
0
CDCl3) d ¼ 16.7 (d, J ¼ 6.0 Hz, POCC), 27.1 (d, J ¼ 140.6 Hz, PC),
50.5 (s, OCH2CH2), 61.9 (d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.5 (d, J ¼ 2.9 Hz,
PCC), 69.3, 112.7 (d, J ¼ 23.0 Hz), 114.9 (d, J ¼ 21.1 Hz), 121.4
(d, J ¼ 2.9 Hz), 121.9, 130.5 (d, J ¼ 8.4 Hz), 133.1 (d, J ¼ 8.4 Hz),
146.7 (d, J ¼ 2.9 Hz), 163.2 (d, J ¼ 244.7 Hz); 31P NMR (243 MHz,
CDCl3) d: 28.46; Anal. calcd. for C15H21FN3O4P: C, 50.42; H, 5.92;
N, 11.76. Found: C, 50.28; H, 6.08; N, 11.49.
–
2 ꢄ POCH2CH3), 1.91 (d, J ¼ 1.2 Hz, 3H, HC CCH ), 2.09
–
3
(dt, J ¼ 18.8 Hz, J ¼ 7.3 Hz, 2H, PCH2), 3.69 (dt, J ¼ 11.7 Hz,
J ¼ 7.3 Hz, 2H, PCH2CH2), 3.82 (t, J ¼ 4.8 Hz, 2H, OCH2CH2),
4.04–4.15 (m, 4H, 2 ꢄ POCH2CH3), 4.53 (t, J ¼ 4.8 Hz, 2H,
Diethyl {2-[4-(2,4-difluorophenyl)-1H-1,2,3-triazol-1-yl]-
ethoxy}methylphosphonate 19c
According to the general procedure (method A) from the azide 13
(0.150 g, 0.630 mmol) and ethynyl-2,4-difluorobenzene 16c
(0.087 g, 0.630 mmol) phosphonate 19c (0.185 g, 78%) was
obtained after chromatography on a silica gel column with
chloroform–methanol (100:1 v/v) as a colorless oil.
–
–
OCH CH ), 4.96 (s, 2H, CH ), 7.35 (d, J ¼ 1.2 Hz, 1H, HC CCH ),
2
2
2
3
7.87 (s, 1H, HC5 ), 8.74 (brs, 1H, NH); 13C NMR (75.5 MHz, CDCl3)
d: 12.5, 16.6 (d, J ¼ 6.1 Hz, POCC), 26.9 (d, J ¼ 139.8 Hz, PC), 43.0,
0
–
50.4, 61.9 (d, J ¼ 6.4 Hz, 2 ꢄ POC), 65.4, 68.9, 111.1, 124.8 (s, HC C),
–
140.2, 142.0 (s, HC C), 151.0 (s, C O), 164.4 (s, C O); 31P NMR
–
–
–
–
–
–
(121.5 MHz, CDCl3) d: 28.29; Anal. calcd. for C16H26N5O6P: C,
46.26; H, 6.31; N, 16.86. Found: C, 45.98; H, 6.18; N, 16.68.
According to the general procedure (method B) from the azide 13
(0.120 g, 0.506 mmol) and ethynyl-2,4-difluorobenzene 16c
(0.070 g, 0.506 mmol) phosphonate 19c (0.163 g, 86%) was obtained
after chromatography on a silica gel column with chloroform–
methanol (100:1 v/v) as a colorless oil; IR (film, cmꢀ1) nmax: 3146,
3102, 2984, 2909, 1493, 1242, 1028, 957, 849, 814; 1H NMR
(600 MHz, CDCl3) d: 1.31 (t, J ¼ 7.1 Hz, 6H, 2 ꢄ POCH2CH3), 3.82
(d, J ¼ 8.4 Hz, 2H, PCH2), 4.05 (t, J ¼ 5.0 Hz, 2H, OCH2CH2), 4.09–
4.16 (m, 4H, 2 ꢄ POCH2CH3), 4.66 (t, J ¼ 5.0 Hz, 2H, OCH2CH2),
6.92 (ddd, J ¼ 11.0 Hz, J ¼ 8.6 Hz, J ¼ 2.5 Hz, 1H, Haromat), 7.00–
Diethyl {2-[4-((2,4-dioxopyrimidin-1-yl)methyl)-1H-1,2,3-
triazol-1-yl]ethoxy}ethylphosphonate 18c
According to the general procedure (method B) from the azide 14
(0.100 g, 0.398 mmol) and N1-propargyluracil 15c (0.060 g,
0.398 mmol) a phosphonate 18c (0.144 g, 90%) was obtained after
chromatography on a silica gel column with chloroform–methanol
(10:1 v/v) as a colorless oil; IR (film, cmꢀ1) nmax: 3152, 2991, 2875,
2823, 1688, 1459, 1239, 1051, 1029, 755; 1H NMR (300 MHz, CDCl3)
d: 1.31 (t, J ¼ 7.0 Hz, 6H, 2 ꢄ POCH2CH3), 2.08 (dt, J ¼ 18.8 Hz,
J ¼ 7.3 Hz, 2H, PCH2), 3.69 (dt, J ¼ 12.2 Hz, J ¼ 7.3 Hz, 2H,
PCH2CH2), 3.82 (t, J ¼ 5.1 Hz, 2H, OCH2CH2), 4.04–4.14 (m, 4H,
2 ꢄ POCH2CH3), 4.53 (t, J ¼ 5.1 Hz, 2H, OCH2CH2), 4.99 (s, 2H,
0
7.05 (m, 1H, Haromat), 8.07 (d, J ¼ 3.8 Hz, 1H, HC5 ); 8.27
(dt, J ¼ 6.5 Hz, J ¼ 8.6 Hz, 1H, Haromat); 13C NMR (151 MHz,
CDCl3) d: 16.3 (d, J ¼ 6.0 Hz, POCC), 50.2, 62.5 (d, J ¼ 6.6 Hz,
2 ꢄ POC), 64.4 (d, J ¼ 167.6 Hz, PC), 71.3 (d, J ¼ 10.6 Hz,
OCH2CH2), 104.0 (d, J ¼ 25.7 Hz), 112.0 (dd, J ¼ 21.7 Hz,
J ¼ 3.3 Hz), 115.1 (dd, J ¼ 13.2 Hz, J ¼ 3.3 Hz), 123.4
(d, J ¼ 12.1 Hz), 128.7 (dd, J ¼ 9.7 Hz, J ¼ 5.2 Hz), 140.4
–
CH ), 5.71 (d, J ¼ 7.9 Hz, 1H, HC CH), 7.53 (d, J ¼ 7.9 Hz, 1H,
–
2
–
–
HC CH), 7.91 (s, 1H, HC ), 9.25 (brs, 1H, NH); 13C NMR (75.5 MHz,
0
5
–
(d, J ¼ 2.3 Hz, HC C), 159.1 (dd, J ¼ 249.9 Hz, J ¼ 10.8 Hz),
CDCl3) d: 16.7 (d, J ¼ 6.0 Hz, POCC), 27.0 (d, J ¼ 140.0 Hz, PC), 43.2,
–
162.3 (dd, J ¼ 249.9 Hz, J ¼ 12.2 Hz); 31P NMR (243 MHz, CDCl3)
d: 20.19; Anal. calcd. for C15H20F2N3O4P: C, 48.00; H, 5.37; N, 11.20.
Found: C, 48.22; H, 5.54; N, 10.96.
–
50.5, 62.2 (d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.4, 68.9, 102.7, 125.0 (s, HC C),
–
141.7 (s, HC C), 144.4, 151.0 (s, C O), 163.9 (s, C O); 31P NMR
–
–
–
–
–
–
(121.5 MHz, CDCl3) d: 28.44; Anal. calcd. for C15H24N5O6P: C,
44.89; H, 6.03; N, 17.45. Found: C, 45.08; H, 6.18; N, 17.49.
Diethyl {2-[4-((6-amino-9H-purin-9-yl)methyl)-1H-1,2,3-
triazol-1-yl]ethoxy}ethylphosphonate 18a
According to the general procedure (method B) from the azide 14
(0.100 g, 0.398 mmol) and N9-propargyladenine 15a (0.069 g,
0.398 mmol) a phosphonate 18a (0.119 g, 70%) was obtained after
Diethyl {2-[4-((4-acetylamino-2-oxopyrimidin-1-yl)methyl)-
1H-1,2,3-triazol-1-yl]ethoxy}ethylphosphonate 18d
According to the general procedure (method B) from the azide 14
(0.100 g, 0.398 mmol) and N4-acetyl-N1-propargylcytosine 15d
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

10
I. E. Głowacka et al.
Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
(0.076 g, 0.398 mmol) a phosphonate 18d (0.125 g, 71%) was
obtained after chromatography on a silica gel column with
Diethyl {2-[4-((1,3-dimethyl-2,6-dioxopurin-7-yl)methyl)-
1H-1,2,3-triazol-1-yl]ethoxy}ethylphosphonate 18g
chloroform–methanol (50:1, 20:1 v/v) as
a colorless oil;
According to the general procedure (method B) from the azide 14
(0.100 g, 0.398 mmol) and N7-propargyltheophylline 15g
(0.087 g, 0.398 mmol) a phosphonate 18g (0.175 g, 94%) was
obtained after chromatography on a silica gel column with
chloroform–methanol (50:1, 20:1 v/v) as a colorless oil; IR
(film, cmꢀ1) nmax: 3111, 2988, 1706, 1661, 1228, 1028, 755;
1H NMR (300 MHz, CDCl3) d: 1.31 (t, J ¼ 7.1 Hz, 6H,
2 ꢄ POCH2CH3), 2.06 (dt, J ¼ 18.7 Hz, J ¼ 7.4 Hz, 2H, PCH2), 3.42
(s, 3H, N–CH3), 3.57 (s, 3H, N–CH3), 3.69 (dt, J ¼ 12.1 Hz, J ¼ 7.4 Hz,
2H, PCH2CH2), 3.81 (t, J ¼ 5.1 Hz, 2H, OCH2CH2), 4.03–4.13 (m, 4H,
2 ꢄ POCH2CH3), 4.52 (t, J ¼ 5.1 Hz, 2H, OCH2CH2), 5.59 (s, 2H, CH2),
IR (film, cmꢀ1) nmax: 3428, 3230, 2981, 2923, 1708, 1665, 1561,
1231, 1051, 795; 1H NMR (300 MHz, CDCl3) d: 1.33 (t, J ¼ 7.1 Hz,
6H, 2 ꢄ POCH2CH3), 2.09 (dt, J ¼ 18.7 Hz, J ¼ 7.5 Hz, 2H, PCH2),
2.23 (s, 3H, C(O)CH3), 3.70 (dt, J ¼ 11.5 Hz, J ¼ 7.5 Hz, 2H,
PCH2CH2), 3.82 (t, J ¼ 4.9 Hz, 2H, OCH2CH2), 4.04–4.14 (m, 4H,
2 ꢄ POCH2CH3), 4.53 (t, J ¼ 4.9 Hz, 2H, OCH2CH2), 5.16 (s, 2H,
–
CH ), 7.37 (d, J ¼ 7.3 Hz, 1H, HC CH), 7.93 (d, J ¼ 7.3 Hz,
–
2
1H, HC CH), 7.95 (s, 1H, HC ), 8.65 (brs, 1H, NH); 13C NMR
–
–
0
5
(75.5 MHz, CDCl3) d: 16.5 (d, J ¼ 6.0 Hz, POCC), 24.9
(s, C(O)CH3), 26.8 (d, J ¼ 139.7 Hz, PC), 43.1, 50.3 (s, OCH2CH2),
62.2 (d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.2, 68.8 (s, OCH2CH2), 97.1,
7.82 (s, 1H), 7.97 (s, 1H, HC5 ); 13C NMR (75.5 MHz, CDCl3) d: 16.7
0
–
–
–
–
125.0 (s, HC C), 141.6 (s, HC C), 148.9, 155.7, 160.3 (s, C O),
–
–
(d, J ¼ 6.1 Hz, POCC), 26.8 (d, J ¼ 140.1 Hz, PC), 28.3, 30.1, 41.7, 50.5,
171.1 (s, C O); 31P NMR (121.5 MHz, CDCl3) d: 28.30; Anal. calcd.
–
–
–
61.8 (d, J ¼ 6.0 Hz, 2 ꢄ POC), 65.4, 68.9, 106.7, 124.9 (s, HC C), 141.6
–
for C17H27N6O6P: C, 46.15; H, 6.15; N, 19.00. Found: C, 46.28;
H, 6.28; N, 18.78.
(s, HC C), 142.1, 148.9, 151.7, 155.4; 31P NMR (121.5 MHz, CDCl3)
d: 28.92; Anal. calcd. for C18H28N7O6P: C, 46.05; H, 6.01; N, 20.89.
Found: C, 45.88; H, 5.83; N, 20.70.
–
–
Diethyl {2-[4-((3,5-dioxo-1,2,4-triazin-2-yl)methyl)-1H-
1,2,3-triazol-1-yl]ethoxy}ethylphosphonate 18e
Diethyl {2-[4-((8-chloro1,3-dimethyl-2,6-dioxopurin-7-yl)-
According to the general procedure (method B) from the azide 14
(0.103 g, 0.410 mmol) and N1-propargyl-6-azauracil 15e (0.062 g,
0.410 mmol) a phosphonate 18e (0.101 g, 62%) was obtained after
chromatography on a silica gel column with chloroform–
methanol (50:1, 20:1 v/v) as a white solid, m.p.: 97–988C; IR
(KBr, cmꢀ1) nmax: 3225, 3110, 3021, 2908, 1730, 1677, 1433,
methyl)-1H-1,2,3-triazol-1-yl]ethoxy}ethylphosphonate 18h
According to the general procedure (method B) from the azide 14
(0.100 g, 0.398 mmol) and 8-chloro-N7-propargyltheophylline
15h (0.101 g, 0.398 mmol) a phosphonate 18h (0.165 g, 83%)
was obtained after chromatography on a silica gel column
with chloroform–methanol (50:1, 20:1 v/v) as a colorless oil;
IR (film, cmꢀ1) nmax: 3144, 2986, 2907, 1707, 1664, 1217, 1049,
757; 1H NMR (300 MHz, CDCl3) d: 1.30 (t, J ¼ 7.0 Hz, 6H,
2 ꢄ POCH2CH3), 2.06 (dt, J ¼ 18.3 Hz, J ¼ 7.0 Hz, 2H, PCH2),
3.41 (s, 3H, N–CH3), 3.54 (s, 3H, N–CH3), 3.68 (dt, J ¼ 12.0 Hz,
J ¼ 5.1 Hz, 2H, PCH2CH2), 3.81 (t, J ¼ 5.1 Hz, 2H, OCH2CH2), 4.03–
4.13 (m, 4H, 2 ꢄ POCH2CH3), 4.51 (t, J ¼ 5.1 Hz, 2H, OCH2CH2),
1
1222, 1027, 756; H NMR (300 MHz, CDCl3) d: 1.34 (t, J ¼ 7.1 Hz,
6H, 2 ꢄ POCH2CH3), 2.10 (dt, J ¼ 18.8 Hz, J ¼ 7.3 Hz, 2H, PCH2),
3.66 (dt, J ¼ 12.1 Hz, J ¼ 7.3 Hz, 2H, PCH2CH2), 3.79 (t, J ¼ 5.0 Hz,
2H, OCH2CH2), 4.03–4.14 (m, 4H, 2 ꢄ POCH2CH3), 4.50
(t, J ¼ 5.0 Hz, 2H, OCH2CH2), 5.21 (s, 2H, CH2), 7.40 (s, 1H), 7.84
(s, 1H, HC5 ), 11.40 (brs, 1H, NH); 13C NMR (75.5 MHz, CDCl3) d: 16.6
0
(d, J ¼ 6.0 Hz, POCC), 26.8 (d, J ¼ 139.9 Hz, PC), 34.7, 50.2, 62.0
5.64 (s, 2H, CH2), 7.92 (s, 1H, HC5 ); 13C NMR (75.5 MHz, CDCl3)
0
–
(d, J ¼ 6.0 Hz, 2 ꢄ POC), 65.1, 68.9, 124.9 (s, HC C), 134.7, 141.5
–
(s, HC C), 149.0 (s, C O), 155.9 (s, C O); 31P NMR (121.5 MHz,
d: 16.5 (d, J ¼ 6.0 Hz, POCC), 26.8 (d, J ¼ 140.3 Hz, PC), 28.1, 29.9,
–
–
–
–
–
–
41.0, 50.2, 61.7 (d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.1, 68.8, 107.3, 124.6
CDCl3) d: 29.42; Anal. calcd. for C14H23N6O6P: C, 41.79; H, 5.76;
N, 20.89. Found: C, 41.58; H, 5.87; N, 21.04.
(s, HC C), 138.8, 141.4 (s, HC C), 147.1, 151.0, 154.4; 31P NMR
–
–
–
–
(121.5 MHz, CDCl3) d: 28.26; Anal. calcd. for C18H27ClN7O6P: C,
42.91; H, 5.40; N, 19.46. Found: C, 43.18; H, 5.36; N, 19.58.
Diethyl {2-[4-((3,7-dimethyl-2,6-dioxopurin-1-yl)methyl)-
1H-1,2,3-triazol-1-yl]ethoxy}ethylphosphonate 18f
According to the general procedure (method B) from the azide
14 (0.100 g, 0.398 mmol) and N1-propargyltheobromine 15f
(0.087 g, 0.398 mmol) a phosphonate 18f (0.172 g, 92%) was
Diethyl {2-[4-((5,6-dimethyl-1H-benzo[d]imidazol-yl)-
methyl)-1H-1,2,3-triazol-1-yl]ethoxy}ethylphosphonate 18i
According to the general procedure (method B) from the azide 14
(0.100 g, 0.398 mmol) and 5,6-dimethyl-N1-propargylbenzimida-
zole 15i (0.073 g, 0.398 mmol) a phosphonate 18i (0.107 g, 62%)
was obtained after chromatography on a silica gel column
with chloroform–methanol (50:1, 20:1 v/v) as a yellow oil; IR
(film, cmꢀ1) nmax: 3114, 2984, 2909, 1224, 1051, 755; 1H NMR
(300 MHz, CDCl3) d: 1.28 (t, J ¼ 7.0 Hz, 6H, 2 ꢄ POCH2CH3), 2.04
(dt, J ¼ 18.7 Hz, J ¼ 7.0 Hz, 2H, PCH2), 2.37 (s, 3H, CH3), 3.38
(s, 3H, CH3), 3.69 (dt, J ¼ 13.4 Hz, J ¼ 6.7 Hz, 2H, PCH2CH2),
3.76 (t, J ¼ 4.8 Hz, 2H, OCH2CH2), 4.02–4.10 (m, 4H,
2 ꢄ POCH2CH3), 4.48 (t, J ¼ 4.8 Hz, 2H, OCH2CH2), 5.45 (s, 2H,
obtained after chromatography on
with chloroform–methanol (50:1, 20:1 v/v) as
a
silica gel column
white
a
solid, m.p.: 57–588C; IR (KBr, cmꢀ1) nmax: 3113, 2986, 2907,
1708, 1663, 1235, 1027, 754; 1H NMR (300 MHz, CDCl3)
d: 1.30 (t, J ¼ 7.2 Hz, 6H, 2 ꢄ POCH2CH3), 2.04 (dt, J ¼ 18.6 Hz,
J ¼ 7.2 Hz, 2H, PCH2), 3.56 (s, 3H, N–CH3), 3.66 (dt, J ¼ 14.7 Hz,
J ¼ 7.2 Hz, 2H, PCH2CH2), 3.79 (t, J ¼ 4.8 Hz, 2H, OCH2CH2),
3.99 (s, 3H, N–CH3), 4.02–4.12 (m, 4H, 2 ꢄ POCH2CH3), 4.48
(t, J ¼ 4.8 Hz, 2H, OCH2CH2), 5.31 (s, 2H, CH2), 7.50 (s, 1H),
7.74 (s, 1H, HC5 ); 13C NMR (75.5 MHz, CDCl3) d: 16.4
0
CH2), 7.82 (s, 1H), 7.97 (s, 1H, HC5 ); 13C NMR (75.5 MHz, CDCl3)
0
(d, J ¼ 6.0 Hz, POCC), 26.6 (d, J ¼ 140.5 Hz, PC), 29.7, 33.6,
35.9, 50.0, 61.7 (d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.0, 68.9, 107.4,
d: 16.5 (d, J ¼ 6.0 Hz, POCC), 20.3, 20.7, 26.8 (d, J ¼ 140.6 Hz, PC),
41.5, 50.3, 61.7 (d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.2 (d, J ¼ 2.0 Hz), 68.8,
110.1, 123.5, 131.1, 132.3, 142.2, 143.0; 31P NMR (121.5 MHz,
CDCl3) d: 28.30; Anal. calcd. for C20H30N5O4P: C, 55.16; H, 6.94;
N, 16.08. Found: C, 55.38; H, 7.13; N, 16.20.
–
–
–
124.3 (s, HC C), 141.6 (s, HC C), 143.2, 148.6, 151.1,
–
154.5; 31P NMR (121.5 MHz, CDCl3) d: 28.06; Anal. calcd.
for C18H28N7O6P: C, 46.05; H, 6.01; N, 20.89. Found: C, 45.92;
H, 6.20; N, 21.09.
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
Synthesis and Biological Evaluation of 1,2,3-Triazolonucleotides
11
(121.5 MHz, CDCl3) d: 28.94; Anal. calcd. for C16H25N4O5P: C, 50.00;
H, 6.56; N, 14.58. Found: C, 49.80; H, 6.37; N, 14.66.
Diethyl {2-[4-(morpholinomethyl)-1H-1,2,3-triazol-1-yl]-
ethoxy}ethylphosphonate 18j
According to the general procedure (method B) from the azide 14
(0.130 g, 0.517 mmol) and N1-propargylmorpholine 15j (0.065 g,
0.517 mmol) a phosphonate 18j (0.114 g, 58%) was obtained after
chromatography on a silica gel column with chloroform–meth-
anol (50:1, 20:1 v/v) as a yellowish oil; IR (film, cmꢀ1) nmax: 2924,
2827, 1210, 1041; 1H NMR (300 MHz, CDCl3) d: 1.32 (t, J ¼ 7.1 Hz,
6H, 2 ꢄ POCH2CH3), 2.07 (dt, J ¼ 18.7 Hz, J ¼ 7.4 Hz, 2H, PCH2),
2.53 (brt, J ¼ 4.6 Hz, 4H, CH2NCH2), 3.67 (s, 2H, CH2), 3.63–3.73
(m, 6H, CH2OCH2, PCH2CH2), 3.83 (t, J ¼ 5.0 Hz, 2H, OCH2CH2),
4.02–4.14 (m, 4H 2 ꢄ POCH2CH3), 4.53 (t, J ¼ 5.1 Hz, 2H,
Diethyl {2-[4-(2-fluorophenyl)-1H-1,2,3-triazol-1-yl]-
ethoxy}ethylphosphonate 20a
According to the general procedure (method B) from the azide 14
(0.114 g, 0.453 mmol) and ethynyl-2-fluorobenzene 16a
(0.055 g, 0.453 mmol) a phosphonate 20a (0.131 g, 78%) was
obtained after chromatography on a silica gel column with
chloroform–methanol (50:1, 20:1 v/v) as a colorless oil; IR
(film, cmꢀ1) nmax: 3147, 2983, 2927, 2875, 1487, 1231, 1048,
963, 818, 764; 1H NMR (300 MHz, CDCl3) d: 1.21 (t, J ¼ 7.1 Hz,
6H, 2 ꢄ POCH2CH3), 2.00 (dt, J ¼ 18.7 Hz, J ¼ 7.3 Hz, 2H, PCH2),
3.64 (dt, J ¼ 12.2 Hz, J ¼ 7.3 Hz, 2H, PCH2CH2), 3.81 (t, J ¼ 5.0 Hz,
2H, OCH2CH2), 3.93–4.05 (m, 4H, 2 ꢄ POCH2CH3), 4.53
(t, J ¼ 5.0 Hz, 2H, OCH2CH2), 7.02–7.24 (m, 3H, Haromat), 8.00
OCH2CH2), 7.68 (s, 1H, HC5 ); 13C NMR (75.5 MHz, CDCl3) d:
0
16.6 (d, J ¼ 6.3 Hz, POCC), 26.9 (d, J ¼ 140.2 Hz, PC), 50.2 (s,
OCH2CH2), 53.4, 53.6, 61.8 (d, J ¼ 6.4 Hz, 2 ꢄ POC), 65.2, 66.8,
69.0, 123.9 (s, HC C), 143.6 (s, HC C); 31P NMR (121.5 MHz, CDCl3)
–
–
–
–
d: 29.05; Anal. calcd. for C15H29N4O5P: C, 47.87; H, 7.77; N, 14.89.
Found: C, 48.00; H, 7.95; N, 14.69.
0
(d, J ¼ 3.8 Hz, 1H, HC5 ); 8.21 (dt, J ¼ 7.6 Hz, J ¼ 2.0 Hz,
1H, Haromat); 13C NMR (75.5 MHz, CDCl3) d: 16.5 (d, J ¼ 6.0 Hz,
POCC), 26.9 (d, J ¼ 140.0 Hz, PC), 50.4 (s, OCH2CH2), 61.8
(d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.3, 69.1, 115.6 (d, J ¼ 21.8 Hz), 118.6
(d, J ¼ 12.9 Hz), 124.5 (d, J ¼ 3.1 Hz), 127.6 (d, J ¼ 3.7 Hz), 129.2
(d, J ¼ 8.3 Hz), 140.9 (d, J ¼ 2.6 Hz), 159.1 (d, J ¼ 247.2 Hz);
31P NMR (121.5 MHz, CDCl3) d: 28.29; Anal. calcd. for
C16H23FN3O4P: C, 51.75; H, 6.24; N, 11.32. Found: C, 52.05;
H, 6.44; N, 11.26.
Diethyl {2-[4-((3-acetyl-1H-indol-1-yl)methyl)-1H-1,2,3-
triazol-1-yl]ethoxy}ethylphosphonate 18k
According to the general procedure (method B) from the azide 14
(0.172 g, 0.685 mmol) and 3-acetyl-N1-propargylindole 15k
(0.135 g, 0.685 mmol) a phosphonate 18k (0.260 g, 85%) was
obtained after chromatography on a silica gel column with
chloroform–methanol (50:1, 20:1 v/v) as a yellowish oil; IR
(film, cmꢀ1) nmax: 3456, 3108, 2982, 2908, 1643, 1528, 1229,
1027, 751; 1H NMR (300 MHz, CDCl3) d: 1.27 (t, J ¼ 7.0 Hz, 6H,
2 ꢄ POCH2CH3), 1.97 (dt, J ¼ 18.5 Hz, J ¼ 7.1 Hz, 2H, PCH2), 2.53
(s, 3H, C(O)CH3), 3.65 (dt, J ¼ 14.1 Hz, J ¼ 7.1 Hz, 2H, PCH2CH2),
3.77 (t, J ¼ 4.7 Hz, 2H, OCH2CH2), 3.97–4.07 (m, 4H,
2 ꢄ POCH2CH3), 4.49 (t, J ¼ 4.7 Hz, 2H, OCH2CH2), 5.47 (s, 2H,
CH2), 7.27–7.34 (m, 2H, Haromat), 7.46–7.50 (m, 1H, Haromat), 7.92
Diethyl {2-[4-(3-fluorophenyl)-1H-1,2,3-triazol-1-yl]-
ethoxy}ethylphosphonate 20b
According to the general procedure (method B) from the azide 14
(0.094 g, 0.374 mmol) and ethynyl-3-fluorobenzene 16b
(0.045 g, 0.374 mmol) a phosphonate 20b (0.136 g, 98%) was
obtained after chromatography on a silica gel column with
chloroform–methanol (50:1, 20:1 v/v) as a colorless oil; IR
(film, cmꢀ1) nmax: 3126, 2973, 2933, 2823, 1484, 1226, 1051,
962, 820, 770; 1H NMR (300 MHz, CDCl3) d: 1.29 (t, J ¼ 7.0 Hz,
6H, 2 ꢄ POCH2CH3), 2.09 (dt, J ¼ 18.5 Hz, J ¼ 7.0 Hz, 2H, PCH2),
3.74 (dt, J ¼ 14.0 Hz, J ¼ 7.0 Hz, 2H, PCH2CH2), 3.87 (t, J ¼ 5.0 Hz,
2H, OCH2CH2), 4.03–4.13 (m, 4H, 2 ꢄ POCH2CH3), 4.65
(t, J ¼ 5.0 Hz, 2H, OCH2CH2), 7.04 (dt, J ¼ 10.9 Hz, J ¼ 2.5 Hz,
1H, Haromat), 7.39–7.44 (m, 1H, Haromat), 7.58–7.64
(s, 1H, HC5 ), 8.36–8.41 (m, 1H, Haromat); 13C NMR (75.5 MHz, CDCl3)
0
d: 16.6 (d, J ¼ 6.1 Hz, POCC), 26.9 (d, J ¼ 140.3 Hz, PC), 27.8, 42.5,
50.4, 61.8 (d, J ¼ 6.1 Hz, 2 ꢄ POC), 65.3, 68.9, 110.0, 117.5, 122.7,
122.7, 123.5, 123.8, 126.4, 135.0, 136.6, 142.8, 193.0; 31P NMR
(121.5 MHz, CDCl3) d: 28.28; Anal. calcd. for C21H29N4O5P: C,
56.24; H, 6.52; N, 12.49. Found: C, 56.56; H, 6.77; N, 12.56.
(m, 2H, Haromat), 8.08 (s, 1H, HC5 ); 13C NMR (75.5 MHz, CDCl3)
0
Diethyl {2-[4-((2-oxopyridin-1-yl)methyl)-1H-1,2,3-triazol-
1-yl]ethoxy}ethylphosphonate 18l
d: 16.4 (d, J ¼ 5.7 Hz, POCC), 50.2 (s, OCH2CH2), 62.5 (d, J ¼ 6.5 Hz,
2 ꢄ POC), 65.3 (d, J ¼ 167.1 Hz, PC), 112.7 (d, J ¼ 23.1 Hz), 114.9
(d, J ¼ 21.1 Hz), 121.2 (d, J ¼ 3.0 Hz), 121.4, 130.4 (d, J ¼ 8.6 Hz),
132.9 (d, J ¼ 8.6 Hz), 146.7 (d, J ¼ 2.7 Hz), 163.2 (d, J ¼ 245.7 Hz);
31P NMR (243 MHz, CDCl3) d: 20.36; Anal. calcd. for
C16H23FN3O4P: C, 51.75; H, 6.24; N, 11.32. Found: C, 51.58;
H, 6.29; N, 11.25.
According to the general procedure (method B) from the azide 14
(0.112 g, 0.446 mmol) and N-propargyl-2-pyridone 15l (0.059 g,
0.446 mmol) a phosphonate 18l (0.146 g, 85%) was obtained after
chromatography on a silica gel column with chloroform–meth-
anol (50:1, 20:1 v/v) as a brown oil; IR (film, cmꢀ1) nmax: 3409,
3136, 2983, 2878, 1659, 1588, 1245, 1050, 770, 732; 1H NMR
(300 MHz, CDCl3) d: 1.32 (t, J ¼ 7.1 Hz, 6H, 2 ꢄ POCH2CH3), 2.07
(dt, J ¼ 18.8 Hz, J ¼ 7.3 Hz, 2H, PCH2), 3.68 (dt, J ¼ 11.3 Hz,
J ¼ 7.3 Hz, 2H, PCH2CH2), 3.82 (t, J ¼ 5.0 Hz, 2H, OCH2CH2),
4.04–4.14 (m, 4H, 2 ꢄ POCH2CH3), 4.50 (t, J ¼ 5.0 Hz, 2H,
OCH2CH2), 5.20 (s, 2H, CH2), 6.18 (dt, J ¼ 8.1 Hz, J ¼ 6.8 Hz,
1H, Haromat), 6.56 (ddd, J ¼ 9.2 Hz, J ¼ 1.3 Hz, J ¼ 0.7 Hz,
1H, Haromat), 7.32 (ddd, J ¼ 9.2 Hz, J ¼ 6.8 Hz, J ¼ 2.0 Hz,
1H, Haromat), 7.61 (ddd, J ¼ 6.8 Hz, J ¼ 2.0 Hz, J ¼ 0.7 Hz,
Diethyl {2-[4-(2,4-difluorophenyl)-1H-1,2,3-triazol-1-yl]-
ethoxy}ethylphosphonate 20c
According to the general procedure (method B) from the azide 14
(0.117 g, 0.466 mmol) and ethynyl-2,4-difluorobenzene 16c
(0.064 g, 0.466 mmol) phosphonate 20c (0.123 g, 68%) was
obtained after chromatography on a silica gel column with
chloroform–methanol (50:1, 20:1 v/v) as a colorless oil; IR
(film, cmꢀ1) nmax: 3145, 3102, 2983, 2876, 1495, 1262, 1027, 957,
849, 810; 1H NMR (300 MHz, CDCl3) d: 1.30 (t, J ¼ 7.0 Hz, 6H,
2 ꢄ POCH2CH3), 2.08 (dt, J ¼ 18.7 Hz, J ¼ 7.2 Hz, 2H, PCH2), 3.71
(dt, J ¼ 14.8 Hz, J ¼ 7.2 Hz, 2H, PCH2CH2), 3.88 (t, J ¼ 4.9 Hz, 2H,
1H, Haromat), 7.88 (s, 1H, HC5 ); 13C NMR (75.5 MHz, CDCl3)
0
d: 16.5 (d, J ¼ 6.0 Hz, POCC), 26.8 (d, J ¼ 140.0 Hz, PC), 44.4,
50.3, 61.8 (d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.2, 68.8, 106.4, 120.6, 124.8
(s, HC C), 137.7, 139.9, 142.4 (s, HC C), 162.0 (s, C O); 31P NMR
–
–
–
–
–
–
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

12
I. E. Głowacka et al.
Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
OCH2CH2), 4.03–4.14 (m, 4H, 2 ꢄ POCH2CH3), 4.61 (t, J ¼ 4.9 Hz,
2H, OCH2CH2), 6.91 (dddd, J ¼ 11.3 Hz, J ¼ 8.9 Hz, J ¼ 2.6 Hz,
J ¼ 0.2 Hz, 1H, Haromat), 7.00 (dddd, J ¼ 8.9 Hz, J ¼ 7.9 Hz,
of 21c in DMSO, methanol or even water was insufficient to
measure the 13C NMR spectrum; IR (KBr, cmꢀ1) nmax: 3320,
3050, 2996, 2899, 1218, 1016, 960, 833, 810; 1H NMR
(600 MHz, CD3OD) d: 3.82 (d, J ¼ 9.0 Hz, 2H, PCH2), 4.09
(t, J ¼ 5.1 Hz, 2H, OCH2CH2), 4.77 (t, J ¼ 5.1 Hz, 2H, OCH2CH2),
7.14–7.18 (m, 2H, Haromat), 8.12–8.08 (m, 2H, Haromat), 8.49
0
J ¼ 2.6 Hz, J ¼ 1.0 Hz, 1H, Haromat), 8.03 (d, J ¼ 3.8 Hz, 1H, HC5 );
8.27 (dt, J ¼ 15.1 Hz, J ¼ 8.7 Hz, 1H, Haromat); 13C NMR (75.5 MHz,
CDCl3) d: 16.5 (d, J ¼ 6.0 Hz, POCC), 26.9 (d, J ¼ 140.0 Hz, PC),
50.4 (s, OCH2CH2), 61.8 (d, J ¼ 6.3 Hz, 2 ꢄ POC), 65.2, 69.0,
104.0 (d, J ¼ 25.5 Hz), 112.0 (dd, J ¼ 21.5 Hz, J ¼ 3.4 Hz),
115.1 (dd, J ¼ 13.5 Hz, J ¼ 3.4 Hz), 123.3 (d, J ¼ 12.3 Hz),
(d, J ¼ 3.0 Hz, 1H, HC5 ); 31P NMR (243 MHz, CD3OD) d: 18.96;
0
Anal. calcd. for C11H12F2N3O4P ꢁ H2O: C, 39.18; H, 4.18; N, 12.46.
Found: C, 38.92; H, 4.49; N, 12.38.
–
128.6 (dd, J ¼ 9.4 Hz, J ¼ 5.2 Hz), 140.4 (s, HC C), 159.1
–
(dd, J ¼ 249.7 Hz, J ¼ 10.6 Hz), 162.3 (dd, J ¼ 249.7 Hz,
J ¼ 12.4 Hz); 31P NMR (121.5 MHz, CDCl3) d: 28.29; Anal. calcd.
for C16H22F2N3O4P: C, 49.36; H, 5.70; N, 10.79. Found: C, 49.12;
H, 5.48; N, 10.50.
{2-[4-((4-Amino-2-oxopyrimidin-1(2H)-yl)methyl)-1H-
1,2,3-triazol-1-yl]ethoxy}methylphosphonic acid 22d
From 17d (0.170 g, 0.397 mmol) phosphonic acid 22d (0.105 g,
80%) was obtained as a white solid; m.p.: 190–1938C; IR (KBr,
cmꢀ1) nmax: 3450, 3323, 3130, 2960, 29 003, 1680, 1245, 1021,
798; 1H NMR (300 MHz, CD3OD) d: 3.74 (d, J ¼ 8.9 Hz, 2H, PCH2),
3.99 (t, J ¼ 5.1 Hz, 2H, OCH2CH2), 4.64 (t, J ¼ 5.1 Hz, 2H,
OCH2CH2), 5.15 (s, 2H, CH2-Cyt), 6.10 (d, J ¼ 7.8 Hz, 1H,
General procedure for the synthesis of phosphonic acids
21a–c, 22d, 22f, 22h, and 22i
Solutions of diethyl phosphonates 19a–c, 17d, 17f, 17h, and 17i
(1.00 mmol) in CH2Cl2 (3 mL) were treated with bromotrimethyl-
silane (10.0 mmol) at room temperature under argon atmos-
phere. The reaction mixture was protected from light and
stirred at room temperature for 24 h. After concentration to
dryness the residue was co-evaporated with dichloromethane
(5 mL) and ethanol (3 ꢄ 3 mL) to afford the crude phosphonic
acids 21a–c, 22d, 22f, 22h, and 22i which were purified by
crystallization with methanol–diethyl ether.
–
–
–
HC CH), 8.06 (d, J ¼ 7.8 Hz, 1H, HC CH), 8.18 (s, 1H,
–
HC5 ); 13C NMR (75.5 MHz, CD3OD) d: 44.7, 52.2, 67.0
(d, J ¼ 162.3 Hz, PC), 72.0 (d, J ¼ 10.5 Hz, OCH2CH2), 95.1,
0
–
–
–
–
–
127.4 (s, HC C), 141.2 (s, HC C), 148.8, 150.8, 161.3 (s, C O);
–
31P NMR (121.5 MHz, CD3OD) d: 19.95; Anal. calcd. for
C10H15N6O5P: C, 36.37; H, 4.58; N, 25.45. Found: C, 36.09;
H, 4.28; N, 25.20.
{2-[4-((3,7-Dimethyl-2,6-dioxopurin-1-yl)methyl)-1H-1,2,3-
triazol-1-yl]ethoxy}methyl phosphonic acid 22f
{2-[4-(2-Fluorophenyl)-1H-1,2,3-triazol-1-yl]ethoxy}-
methylphosphonic acid 21a
From 17f (0.070 g, 0.15 mmol) phosphonic acid 22f (0.048 g,
78%) was obtained as a white solid; m.p.: 188–1928C; IR (KBr,
cmꢀ1) nmax: 3302, 3110, 2983, 2905, 1717, 1668, 1216, 1024, 970
From 19a (0.082 g, 0.23 mmol) phosphonic acid 21a (0.058 g,
84%) was obtained as a white solid; m.p.: 193–1968C; solubility
of 21a in DMSO, methanol or even water was insuficient to
measure the 13C NMR spectrum; IR (KBr, cmꢀ1) nmax: 3320,
3050, 2993, 2892, 1225, 1024, 980, 810, 769; 1H NMR
(600 MHz, CD3OD) d: 3.73 (d, J ¼ 8.6 Hz, 2H, PCH2), 4.00
(t, J ¼ 5.0 Hz, 2H, OCH2CH2), 4.67 (t, J ¼ 5.0 Hz, 2H, OCH2CH2),
7.13 (brt, J ¼ 9.9 Hz, 1H, Haromat), 7.19 (dt, J ¼ 7.3 Hz,
1H, Haromat), 7.31–7.34 (m, 1H, Haromat), 8.32 (brt, J ¼ 7.3 Hz,
cmꢀ1 1H NMR (600 MHz, CD3OD) d: 3.57 (s, 3H, N–CH3), 3.76
;
(d, J ¼ 8.8 Hz, 2H, PCH2), 4.04 (s, 3H, N–CH3), 4.05 (t, J ¼ 4.9 Hz,
2H, OCH2CH2), 4.77 (t, J ¼ 4.9 Hz, 2H, OCH2CH2), 5.38 (s, 2H, CH2),
8.15 (s, 1H), 8.45 (s, 1H); 13C NMR (151 MHz, D2O) d: 30.4, 34.2,
35.4, 51.1, 65.4 (d, J ¼ 159.8 Hz, PC), 70.6 (d, J ¼ 11.8 Hz,
–
–
–
OCH CH ), 107.7, 126.5 (s, HC C), 141.6 (s, HC C), 142.2, 146.6,
–
2
2
151.6, 155.0; 31P NMR (243 MHz, CD3OD) d: 18.59; Anal. calcd.
for C13H18N7O6P ꢁ H2O: C, 37.42; H, 4.83; N, 23.49. Found: C,
37.68; H, 4.70; N, 23.24.
1H, Haromat); 8.23 (brs,1H, HC5 ); 31P NMR (243 MHz, CD3OD)
0
d: 18.95; Anal. calcd. for C11H13FN3O4P ꢁ 2H2O: C, 39.18;
H, 5.08; N, 12.46. Found: C, 39.08; H, 5.19; N, 12.40.
{2-[4-((8-Chloro-1,3-dimethyl-2,6-dioxopurin-7-yl)methyl)-
{2-[4-(3-Fluorophenyl)-1H-1,2,3-triazol-1-yl]ethoxy}-
methylphosphonic acid 21b
1H-1,2,3-triazol-1-yl]ethoxy}methylphosphonic acid 22h
From 17h (0.082 g, 0.17 mmol) phosphonic acid 22h (0.065 g,
89%) was obtained as a white solid; m.p.: 204–2078C; solubility
of 22h in DMSO, methanol or even water was insuficient to
measure the 13C NMR spectrum; IR (KBr, cmꢀ1) nmax: 3260,
3004, 2983, 2880, 1668, 1230, 1022, 755; 1H NMR (600 MHz,
CD3OD) d: 3.24 (s, 3H, N–CH3), 3.29 (s, 3H, N–CH3), 3.64
(d J ¼ 8.8 Hz, 2H, PCH2), 3.99 (t, J ¼ 5.6 Hz, 2H, OCH2CH2),
4.57 (t, J ¼ 5.6 Hz, 2H, OCH2CH2), 5.60 (s, 2H, CH2), 8.04 (s, 1H,
From 19b (0.110 g, 0.31 mmol) phosphonic acid 21b (0.074 g,
80%) was obtained as a white solid; m.p.: 187–1928C; solubility of
21b in DMSO, methanol or even water was insuficient to measure
the 13C NMR spectrum; IR (KBr, cmꢀ1) nmax: 3300, 3102, 2989,
2930, 1224, 1025, 960, 820, 772; 1H NMR (600 MHz, CD3OD)
d: 3.74 (d, J ¼ 8.5 Hz, 2H, PCH2), 4.00 (t, J ¼ 5.2 Hz, 2H,
OCH2CH2), 4.61 (t, J ¼ 5.2 Hz, 2H, OCH2CH2), 7.05–7.08
HC5 ); 31P NMR (243 MHz, CD3OD) d: 19.14; Anal. calcd. for
0
0
(m, 1H, Haromat), 7.35–7.45 (m, 3H, Haromat), 8.28 (s, 1H, HC5 );
31P NMR (121.5 MHz, CD3OD) d: 18.77; Anal. calcd.
for C11H13FN3O4P ꢁ H2O: C, 41.39; H, 4.74; N, 13.16. Found: C,
41.16; H, 4.51; N, 13.44.
C13H17ClN7O6P: C, 36.00; H, 3.95; N, 22.13. Found: C, 36.28;
H, 4.08; N, 22.28.
{2-[4-((5,6-Dimethyl-1H-benzo[d]imidazol-yl)methyl)-1H-
1,2,3-triazol-1-yl]ethoxy}methylphosphonic acid 22i
{2-[4-(2,4-Difluorophenyl)-1H-1,2,3-triazol-1-yl]ethoxy}-
methylphosphonic acid 21c
From 19c (0.104 g, 0.28 mmol) phosphonic acid 21c (0.055 g,
63%) was obtained as a white solid; m.p.: 180–1838C; solubility
From 17i (0.055 g, 0.13 mmol) phosphonic acid 22i (0.040g, 82%)
was obtained as a white solid; m.p.: 157–1598C; IR (KBr, cmꢀ1
)
nmax: 3335, 3090, 2980, 2931, 1225, 1039, 971, 835; 1H NMR
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
Synthesis and Biological Evaluation of 1,2,3-Triazolonucleotides
13
(600 MHz, CD3OD) d: 2.48 (s, 3H, CH3), 2.50 (s, 3H, CH3), 3.76
(d, J ¼ 8.9 Hz, 2H, PCH2), 4.00 (t, J ¼ 5.0 Hz, 2H, OCH2CH2),
4.66 (t, J ¼ 5.0 Hz, 2H, OCH2CH2), 5.84 (s, 2H, CH2-IMID), 7.63
(s, 1H), 7.80 (s, 1H), 8.35 (s, 1H), 9.40 (s, 1H); 13C NMR (151 MHz,
CD3OD) d: 19.1, 19.3, 41.6, 50.2, 65.8 (d, J ¼ 166.1 Hz, PC), 70.8
[2] C. K. Chu, Antiviral Nucleosides: Chiral Synthesis and
Chemotherapy, Elsevier B.V., Amsterdam 2003.
[3] P. Merino, Curr. Med. Chem. 2006, 13, 539–545.
[4] Y.-S. Lee, B. H. Kim, Bioorg. Med. Chem. Lett. 2002, 12, 1395–
1397.
–
(d, J ¼ 12.1 Hz, OCH CH ), 112.7, 114.0, 125.7 (s, HC C), 129.6,
–
2
2
129.4, 137.1, 137.4, 139.6 (s, HC C), 139.9; 31P NMR (243 MHz,
[5] S. Pan, N. M. Amankulor, K. Zhao, Tetrahedron 1998, 54, 6587–
6604.
–
–
CD3OD) d: 18.74; Anal. calcd. for C15H20N5O4P ꢁ H2O: C, 47.00;
[6] V. Mannucci, F. M. Cordero, A. Piperno, G. Romeo, A. Brandi,
Tetrahedron: Asymmetry 2008, 19, 1204–1209.
H, 5.78; N, 18.27. Found: C, 46.88; H, 5.64; N, 18.45.
´
[7] G. Romero, S. V. Giofre, A. Piperno, R. Romero, M. A.
Chiacchio, Arkivoc 2009 (viii), 168–176.
Antiviral activity assays
The antiviral assays were based on inhibition of virus-induced
cytopathicity in HEL [herpes simplex virus type 1 (HSV-1), HSV-2
(G), vaccinia virus and vesicular stomatitis virus], Vero (para-
influenza-3, reovirus-1, Sindbis, Coxsackie B4, and Punta Toro
virus), HeLa (vesicular stomatitis virus, Coxsackie virus B4, and
respiratory syncytial virus), MDCK (influenza A (H1N1 and
H3N1) and influenza B virus), or CRFK (feline herpes virus;
FIPV) cell cultures. Confluent cell cultures in microtiter
96-well plates were inoculated with 100 cell culture inhibitory
dose-50 (CCID50) of virus (1 CCID50 being the virus dose to
infect 50% of the cell cultures) in the presence of varying con-
centrations (250, 50, 10, . . . mM) of the test compounds.
Viral cytopathicity was recorded as soon as it reached
completion in the control virus-infected cell cultures that
were not treated with the test compounds. The antiviral concen-
tration was expressed as the EC50 or 50%-effective compound
concentration required to inhibit virus-induced cytopathicity
by 50%.
[8] U. Chiacchio, F. Genovese, D. Iannazzo, V. Librando, P.
Merino, A. Rescifina, R. Romero, A. Procopio, G. Romero,
Tetrahedron 2004, 60, 441–448.
[9] U. Chiacchio, M. G. Saita, L. Crispino, G. Gumina,
S. Mangiafico, V. Pistara, G. Romero, A. Piperno, E. De Clercq,
Tetrahedron 2006, 62, 1171–1181.
[10] H. B. Lazrek, M. Taourirte, T. Oulih, J. L. Barascut, J. L.
Imbach, C. Pannecouque, M. Witvrouw, E. De Clercq,
Nucleosides, Nucleotides, Nucleic Acids 2001, 20, 1949–1960.
[11] O. Moukha-Chafiq, M. L. Taha, H. B. Lazrek, C. Pannecouque,
M. Witvrouw, E. De Clercq, Nucleosides, Nucleotides, Nucleic
Acids 2001, 20, 1811–1821.
[12] O. Moukha-Chafiq, M. L. Taha, H. B. Lazrek, J. J. Vasseur,
C. Pannecouque, M. Witvrouw, E. De Clercq, J. L. Barascut,
J. L. Imbach, Nucleosides, Nucleotides, Nucleic Acids 2001, 20,
1797–1810.
[13] J. Krim, M. Taourirte, J. W. Engels, Molecules 2012, 17, 179–190.
[14] S. Trakossas, E. Coutouli-Argyopoulou, D. Hadjipavlou-
Litina, Tetrahedron Lett. 2011, 14, 1673–1676.
Cytotoxicity and cytostatic assay
The cytotoxicity of the test compounds was monitored as a
microscopically visible alteration of cell morphology, and
expressed as the minimal cytotoxic concentration (MCC) or
compound concentration required to afford a microscopically
detectable alteration of cell culture morphology.
The cytostatic activity of the test compounds was determined
as the 50% cytostatic concentration (IC50) or compound concen-
tration required to inhibit cell proliferation by 50%. For this
purpose, cells were seeded in 200 ml-wells of 96-well microtiter
plates and allowed to proliferate for 2 (L1210) to 3 (CEM, HeLa)
days in the absence or presence of different serial concentrations
of the test compounds. At the end of the exponential prolifer-
ation phase, the cells were counted by an automated Coulter ZI
particle counter (Analis, Ghent, Belgium).
[15] H. Elayadi, M. Smietana, Ch. Pannecouque, P. Leyssen, J.
Neyts, J.-J. Vasseur, H. B. Lazrek, Bioorg. Med. Chem. Lett. 2010,
20, 7365–7368.
[16] K. Ishiyama, G. E. Smyth, T. Ueda, Y. Masutomi, T. Oghi, J.
Yano, J. Am. Chem. Soc. 2004, 126, 7476–7485.
[17] N. Hossian, N. Blaton, O. Peeters, J. Rozenski, P. A.
Herdewijn, Tetrahedron 1996, 52, 5563–7778.
[18] M. W. Chun, J. H. Kim, M. J. Kim, B. R. Kim, Nucleosides,
Nucleotides, Nucleic Acids 2001, 24, 979–981.
[19] H. B. Lazrek, M. Taourirte, T. Oulih, Y. Kabbaj, J. L. Barascut,
J. L. Imbach, N. A. Almasoudi, W. Pfleiderer, Nucleosides
Nucleotides 1997, 16, 1073–1077.
[20] P. Chittepu, V. R. Sirivolu, F. Seela, Bioorg. Med. Chem. 2008,
16, 8427–8439.
The authors wish to express their gratitude to Mrs. Małgorzata Pluskota,
Mrs. Leentje Persoons, Mrs. Frieda De Meyer, and Mrs. Lizette van
Berckelaer for excellent technical assistance. This work was supported
by the National Science Centre under Decision UMO-2011/01/D/NZ4/
01276. The virological part of this work was supported by the K.U.
Leuven (GOA no. 10/014).
[21] J.-L. Yu, Q.-P. Wu, Q.-S. Zhang, Y.-H. Liu, Y.-Z. Li, Z.-M. Zhou,
Bioorg. Med. Chem. Lett. 2010, 20, 240–243.
[22] D. G. Piotrowska, J. Balzarini, I. E. Głowacka, Eur. J. Med. Chem.
2012, 47, 501–509.
[23] M. Aufort, J. Herscovici, P. Bouhours, N. Moreau, C. Girard,
Bioorg. Med. Chem. Lett. 2008, 18, 1195–1198.
The authors have declared no conflict of interest.
[24] J. A. Demaray, J. E. Thuener, M. N. Dawson, S. J. Sucheck,
Bioorg. Med. Chem. Lett. 2008, 18, 4868–4871.
[25] X.-L. Wang, K. Wan, Ch.-H. Zhou, Eur. J. Med. Chem. 2010, 45,
4631–4639.
References
[26] D. C. M. Chan, C. A. Laughton, S. F. Queener, M. F. G. Stevens,
Bioorg. Med. Chem. 2002, 10, 3001–3010.
[1] P. Herdewijn, Modified Nucleosides in Biochemistry, Biotechnology
and Medicine, Wiley-VCH, Weinheim 2008.
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

14
I. E. Głowacka et al.
Arch. Pharm. Chem. Life Sci. 2013, 000, 1–14
[27] D. B. Jordan, G. S. Basarab, D.-I. Liao, W. M. P. Johnson, K. N.
Winzenberg, D. A. Winkler, J. Mol. Graphics Modell. 2001, 19,
434–447.
[45] A. Holy`, Curr. Pharm. Des. 2003, 9, 2567–2592.
[46] E. De Clercq, Biochem. Pharm. 2007, 73, 911–922.
´
`
[47] D. T. Keough, D. Hockova, A. Holy, L. M. J. Naesens, T. S.
Skinner-Adams, J. de Jersey, L. W. Guddat, J. Med. Chem. 2009,
52, 4391–4399.
[28] L. V. R. Reddy, P. V. Reddy, N. N. Mishra, P. K. Shukla,
G. Yadav, R. Srivastava, A. K. Shaw, Carbohydr. Res. 2010,
345, 1515–1521.
´
`
´
[48] D. Hockova, A. Holy, M. Masojidkova, D. T. Keough,
J. de Jersey, Bioorg. Med. Chem. 2009, 17, 6218–6232.
[29] A. Kamal, N. Shankaraiah, V. Devaiah, K. L. Reddy,
A. Juvekar, S. Sen, N. Kurian, S. Zingde, Bioorg. Med. Chem.
Lett. 2008, 18, 1468–1473.
[49] I. E. Głowacka, Tetrahedron: Asymmetry 2009, 20, 2270–2278.
´
[50] I. E. Głowacka, M. Cieslak, D. G. Piotrowska, Phosphorus, Sulfur
Silicon 2011, 186, 431–449.
[30] Y. Chen, M. Lopez-Sanchez, D. N. Savoy, D. D. Billadeau,
G. S. Dow, A. P. Kozikowski, J. Med. Chem. 2008, 51, 3437–
3448.
´
[51] A. E. Wroblewski, I. E. Głowacka, Tetrahedron: Asymmetry
2004, 15, 1457–1464.
[31] J. Vantikommu, S. Palle, P. S. Reddy, V. Ramanatham,
M. Khagga, V. R. Pallapothula, Eur. J. Med. Chem. 2010, 45,
5044–5050.
´
[52] A. E. Wroblewski, I. E. Głowacka, Tetrahedron: Asymmetry
2005, 16, 4056–4064.
´
[53] I. E. Głowacka, J. Balzarini, A. E. Wroblewski, Nucleosides,
Nucleotides, Nucleic Acids 2012, 31, 293–318.
[32] K. E. Akri, K. Bougrin, J. Balzarini, A. Faraj, R. Benhida,
Bioorg. Med. Chem. Lett. 2007, 17, 6656–6659.
ˇ´
´
[54] A. Holy`, I. Rosenberg, H. Dvorakova, Collect. Czech. Chem.
Commun. 1989, 54, 2190–2209.
[33] S.-J. Yan, Y.-J. Liu, Y.-L. Chen, L. Liu, J. Lin, Bioorg. Med. Chem.
Lett. 2010, 20, 5225–5228.
ˇ
´
´
´
[55] A. Holy`, I. Votruba, E. Tloust’ova, M. Masojıdkova, Collect.
Czech. Chem. Commun. 2001, 66, 1545–1592.
[34] L. Zhou, A. Amer, M. Korn, R. Burda, J. Balzarini, E.
De Clercq, E. R. Kern, P. F. Torrence, Antiviral Chem.
Chemother. 2005, 16, 375–383.
[56] K. Eger, E. M. Klu¨nder, M. Schmidt, J. Med. Chem. 1994, 37,
3057–3061.
[35] S. Mohan, S. McAtamney, T. Haselhorst, M. von Itzstein,
B. M. Pinto, J. Med. Chem. 2010, 53, 7377–7391.
´
[57] M. Koszytkowska-Stawinska, E. De Clercq, J. Balzarini,
Bioorg. Med. Chem. 2009, 17, 3756–3762.
[36] X. B. Chen, D. Q. Shi, Phosphorus, Sulfur Silicon 2008, 183, 1134–
1144.
´
[58] A. De la Hoz, A. Dıaz-Ortis, A. Moreno, F. Langa, Eur. J. Org.
Chem. 2000, 3659–3673.
[37] J. M. Cox, T. R. Hawkes, P. Bellini, R. M. Ellis, R. Barrett,
J. J. Swanborough, S. E. Russell, P. A. Walker, N. J. Barnes,
A. J. Knee, T. L. Lewis, P. R. Davies, Pestic. Sci. 1997, 50, 297–
311.
[59] Y. Saito, V. Escuret, D. Durantel, F. Zoulim, R. F. Schinazi,
L. A. Agrofoglio, Bioorg. Med. Chem. 2003, 11, 3633–3639.
[60] L. Lolk, J. Pøhlsgaard, A. S. Jepsen, L. H. Hansen, H. Nielsen,
S. I. Steffansen, L. Sparving, A. B. Nielsen, B. Vester, P. Nielsen,
J. Med. Chem. 2008, 51, 4957–4967.
[38] K. Ramalingam, N. Raju, P. Nanjappan, K. E. Linder, J. Pirro,
W. Zeng, W. Rumsey, D. P. Nowotnik, A. D. Nunn, J. Med.
Chem. 1994, 37, 4155–4163.
[61] W. E. Lindsell, Ch. Murray, P. N. Preston, T. A. J. Woodman,
Tetrahedron 2000, 56, 1233–1245.
[39] M. J. Genin, D. A. Allwine, D. J. Anderson, M. R. Barbachyn,
D. E. Emmert, S. A. Garmon, D. R. Graber, K. C. Grega, J. B.
Hester, D. K. Hutchinson, J. Morris, R. J. Reischer, Ch. W.
Ford, G. E. Zurenko, J. C. Hamel, R. D. Schaadt, D. Stapert,
B. H. Yagi, J. Med. Chem. 2000, 43, 953–970.
[62] A. Casaschi, R. Grigg, J. M. Sansano, Tetrahedron 2000, 56,
7553–7560.
[63] M. J. A. Wear, P. A. M. Herdewijn, W. E. Pfleiderer,
Immunosuppresive effect of 8-substitued xanthine deriva-
tives. United States Patent No. US 7,253,176 B1.
[40] P. Norris, D. Horton, B. R. Levine, Heterocycles 1996, 43, 2643–
2655.
[64] J. W. Daly, W. L. Padgett, M. T. Shamim, J. Med. Chem. 1986, 29,
1305–1308.
[41] R. R. Talekar, R. H. Wightman, Tetrahedron 1997, 53, 3831–
3842.
[65] A. Casaschi, R. Grigg, J. M. Sansano, Tetrahedron 2001, 57,
607–615.
[42] C. W. Tornøe, C. Christensen, M. Meldal, J. Org. Chem. 2002,
67, 3057–3064.
[66] J. Verron, P. Malherbe, E. Prinssen, A. W. Thomas, N. Nock,
R. Masciadri, Tetrahedron Lett. 2007, 48, 377–380.
[43] V. V. Rostovtsev, L. G. Green, V. V. Fokin, K. B. Sharpless,
Angew. Chem. Int. Ed. 2002, 41, 2596–2599.
´
´
[67] L. Perez-Serrano, L. Casarrubios, G. Dominguez, P. Gonzalez-
´ ´
Perez, J. Perez-Castells, Synthesis 2002, 13, 1810–1812.
[44] E. De Clercq, A. Holy`, Nat. Rev. Drug Discov. 2005, 4, 928–940.
ß 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com