Synthesis and antiviral evaluation of C5-substituted-(1,3-diyne)-2′- deoxyuridines

Article abstract of DOI:10.1016/j.ejmech.2012.04.001

Starting from acetylated 5-ethynyl-2′-deoxyuridine (3), 14 hitherto unknown C5-substituted-(1,3-diyne)-2′-deoxyuridines (with cyclopropyl, hydroxymethyl, methylcyclopentane, p-(substituted)phenyl and disubstituted-phenyl substituents) have been synthesized via a nickel-copper catalyzed C-H activation between two terminal alkynes, in yields ranging from 19% to 67%. Their antiviral activities were measured against a large number of DNA and RNA viruses including herpes simplex virus type 1 and type 2, varicella-zoster virus, human cytomegalovirus and vaccinia virus. The 5-[4-(4-trifluoromethoxyphenyl)buta-1,3-diynyl]-2′-deoxyuridine (26) is the most potent inhibitor of this series against VZV with an EC₅₀ of ～1 μM and a CC₅₀ of 55 μM. Their cytostatic activities were determined against murine leukemia cells, human T-lymphocyte cells and cervix carcinoma cells. Compounds were also evaluated on a wide panel of RNA viruses, including influenza virus A (H1N1 and H3N2) and B in MDCK cell cultures, parainfluenza-3 virus, reovirus-1, Sindbis virus and Punta Toro virus in Vero cell cultures and vesicular stomatitis, coxsackie B4 and respiratory syncytial virus in HeLa cell cultures and against human immunodeficiency virus type 1 and 2 in CEM cell cultures, with no specific antiviral effect. This class of compounds could be of further interest for lead optimization as anti-infectious (i.e. herpetic) agents.

Full text of DOI:10.1016/j.ejmech.2012.04.001

European Journal of Medicinal Chemistry 53 (2012) 220e228
Contents lists available at SciVerse ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Original article
Synthesis and antiviral evaluation of C5-substituted-(1,3-diyne)-20-deoxyuridines
,
Ozkan Sari a, Vincent Roy a, Jan Balzarini b, Robert Snoeck b, Graciela Andrei b, Luigi A. Agrofoglio a
*
a Institut de Chimie Organique et Analytique, UMR 7311 CNRS, Université d’Orléans, UFR Science-rue de chartres, 45067 Orléans, France
b Rega Institute for Medical Research, KU Leuven, 3000 Leuven, Belgium
a r t i c l e i n f o
a b s t r a c t
Starting from acetylated 5-ethynyl-20-deoxyuridine (3), 14 hitherto unknown C5-substituted-(1,3-diyne)-
20-deoxyuridines (with cyclopropyl, hydroxymethyl, methylcyclopentane, p-(substituted)phenyl and
disubstituted-phenyl substituents) have been synthesized via a nickel-copper catalyzed CeH activation
between two terminal alkynes, in yields ranging from 19% to 67%. Their antiviral activities were measured
against a large number of DNA and RNA viruses including herpes simplex virus type 1 and type 2, vari-
cella-zoster virus, human cytomegalovirus and vaccinia virus. The 5-[4-(4-trifluoromethoxyphenyl)buta-
1,3-diynyl]-20-deoxyuridine (26) is the most potent inhibitor of this series against VZV with an EC50 of
Article history:
Received 8 December 2011
Received in revised form
19 March 2012
Accepted 3 April 2012
Available online 21 April 2012
Keywords:
Diyne scaffold
w1 mM and a CC50 of 55 mM. Their cytostatic activities were determined against murine leukemia cells,
human T-lymphocyte cells and cervix carcinoma cells. Compounds were also evaluated on a wide panel of
RNA viruses, including influenza virus A (H1N1 and H3N2) and B in MDCK cell cultures, parainfluenza-3
virus, reovirus-1, Sindbis virus and Punta Toro virus in Vero cell cultures and vesicular stomatitis, cox-
sackie B4 and respiratory syncytial virus in HeLa cell cultures and against human immunodeficiency virus
type 1 and 2 in CEM cell cultures, with no specific antiviral effect. This class of compounds could be of
further interest for lead optimization as anti-infectious (i.e. herpetic) agents.
Nucleoside
Nickel-copper catalyzed CeH activation
Anti-VZV activity
Ó 2012 Published by Elsevier Masson SAS.
1. Introduction
particular HSV-1 and VZV) [7]. Bicyclic furano pyrimidine nucleo-
sides, pioneered by McGuigan et al. [8] have been found to be highly
C5-substituted pyrimidine nucleosides, especially those in the 20-
deoxyuridine series, form a class of unique compounds because of
the important role they play as components of nucleotide-derived
tools for molecular genetics [1] but also as antiviral and anticancer
agents [2]. For example (Fig. 1), several groups have focused on the
introduction of a fluorescent probe at C5 of 20-deoxyuridine [3,4]
meanwhile others have developed C5-substituents that influence
duplex annealing [5,6]; finally, a great deal of effort has been
expended to develop C5-substituted analogues that can be incor-
porated into DNA by DNA polymerases; for instance, the 5-(2-
substituted vinyl)-20-deoxyuridines (e.g., BVDU, BrivudinÒ) have
emerged as potent and selective inhibitors of herpes viruses (in
potent and selective inhibitors of varicella-zoster virus (VZV) espe-
cially with the presence of a long alkyl or aryl chain at the 6-position
of the furano ring. Those compounds exhibited high lipophilicity
values with a better membrane permeation. Sawai et al. [9] have
described 20-deoxyuridines containing C5-substituents linked
through a C5-carbonylmethylene, C5-alkynyl or C5-alkenyl groups.
More recently, Rai et al. [10] reported novel C5-alkynylpyrimidine
nucleosides as potent inhibitors of mycobacteria, whereas similar
analogues werefound tobe potentinhibitorsof humanbreast cancer
cell proliferation [11]. It appears from those data that the C5 position
of the pyrimidine nucleosides can accept even large groups without
interfering with DNA duplex formation or incorporation into DNA,
even if there is no well-defined ideal combination of the substituent
on the nucleoside and the enzyme. On a chemical point of view, most
approaches to C5-substituted pyrimidine nucleosides are based on
variations of the organo-palladium-mediated Heck, Sonogashira,
Stille or Suzuki reactions between the 5-halopyrimidine moiety to
alkenyl, alkynyl, carbonyl and aryl moieties [12].
Abbreviations: VZV-zoster virus, varicella; VV, vaccinia virus; HSV, herpes
simplex virus; VSV, vesicular stomatitis virus; DNA, deoxyribonucleic acid; RNA,
ribonucleic acid; EC50, compound concentration affording 50% inhibition of the viral
cytopathicity; CC50, compound concentration affording 50% inhibition of cytotox-
icity; MDCK, Madin-Darby canine kidney; MW, microwave; TMEDA, tetramethy-
lenediamine; DMF, dimethylformamide; TBAF, tetrabutylammonium fluoride; THF,
tetrahydrofuran; DMSO, dimethylsulfoxide; EDG, electron donating group; EWG,
electron withdrawing group.
As mentioned above, whereas C5-alkynylpyrimidine nucleosides
are known compounds, less efforts have been done for the devel-
opment of C5-substituted-1,3-diynyl-20-deoxyuridines. However,
1,4-disubstituted 1,3-butadiynes are found in biological molecules
* Corresponding author. Tel.: þ33 2 3849 4582.
E-mail address: luigi.agrofoglio@univ-orleans.fr (L.A. Agrofoglio).
0223-5234/$ e see front matter Ó 2012 Published by Elsevier Masson SAS.
doi:10.1016/j.ejmech.2012.04.001

O. Sari et al. / European Journal of Medicinal Chemistry 53 (2012) 220e228
221
O
O
N
HN
spacer-fluorophore
O
N
O
N
OH
Br
HN
HN
NH
O
O
HO
O
HO
HO
O
O
O
OH
OH
OH
one fluorophore analogue
[3,4]
one C5-substituted analogue
for DNA duplex annealing, [5,6]
BVDU
anti-HSV, [8]
R
R
O
N
R
O
N
O
HN
NH
O
HO
HO
N
O
HO
N
O
O
O
O
18-31
OH
OH
OH
R = aryl, substituted-aryl,
C5-substituted alkynyl analogues
Furanopyrimidine analogues
potent anti-VZV, [9]
pyridine, alkyl,
antimycobacterial [11] or antitumoral [12]
(hydroxy)alkyl, cycloalkyl
Fig. 1. Some C5-substituted pyrimidine nucleosides and target compounds (18e31).
and serve as intermediates in the synthesis of heterocycles of
pharmaceutical interest [13]. To the best of our knowledge only, two
teams, Minakawa et al. [14] and Brown et al. [15] have reported
C5-substituted 1,3-diyne nucleoside derivatives such as the C5-
(4-anthracenylbuta-1,3-diynyl)-20-deoxyuridine, but these nucleo-
sides were obviously meant to be incorporated into oligonucleotides
for their fluorescence properties and none of the C5-(1,3-diyne)
nucleosides have been evaluated for their antiviral properties. Taken
together, all the mentioned studies [3e11] indicate generally the
requirement of a nonpolar moietyat C5 position of a pyrimidine ring
or at C6 of a furano moiety. To probe this requirement in these new
C5-substituted-1,3-diynyl-20-deoxyuridines (18e31), we chose to
incorporate at C5 a series of substituted (alkyl, amino, nitro, .)
phenyl groups and we report herein on the synthesis and charac-
terization of these novel analogues targeting virus replication for
inhibitory activity.
Thus, the 30,50-diacetylated-20-deoxyuridine (1) was first con-
verted to the key intermediate C5-ethynyl analogue (3) following
a pathway already established previously by our team [23]. The
iodination of 1 in the presence of ammonium cerium nitrate in
acetonitrile afforded 2 in 87% yield. The resulting pure acetylated 5-
iodouracil 2 was reacted, under Sonogashira conditions, with tri-
methylsilylacetylene (TMS) in the presence of Et3N, CuI and
PdCl2(PPh3)2 in anhydrous DMF at room temperature to afford after
desilylation the desired 5-ethynyluracil nucleoside 3 in 75% overall
yield.
In a first attempt (Method A), we decided to adapt the Pd/C and
CuI homocoupling reaction, developed by Sajiki et al. [24], which
catalyzed the oxidative dimerization of terminal alkynes and
O
N
O
N
O
N
I
NH
NH
NH
AcO
O
AcO
O
AcO
O
O
O
O
i
ii
2. Results and discussion
2.1. Chemistry
OAc
1
OAc
2
OAc
3
Glaser [16] has pioneered the synthesis of 1,3-diynes through
a Cu(I)-catalyzed oxidative homocoupling of terminal alkynes;
since then, several systems have been developed such as the Hay
coupling [17] based on Cu(I)-TMEDA, the Eglinton reaction [18]
which uses a stoechiometric amount of Cu(II)-salt, or recently the
SonogashiraeGlaser homocoupling sequence reported by Merkul
et al. [19]. Other catalytic systems rely on combined Pd(0) or Pd(II)
with Cu(I) [20,21]. Nevertheless, those reactions are often limited to
the formation of symmetrical diynes. This limitation can be over-
come by the widely used CadioteChodkiewicz [22] reaction which
is actually the Cu-catalyzed coupling of a haloalkyne and a termi-
nal alkyne. Thus, based on the development of various catalytic
iii or
iv
R
R
O
N
O
N
NH
NH
HO
O
AcO
O
v
O
O
OH
OAc
18-31
4-17
Scheme 1. Reagents and conditions: i) I2/CAN, CH3CN, reflux, 87%; ii) (a) trimethysi-
lylacetylene, CuI, PdCl2(PPh3)2, DMF, Et3N; (b) TBAF, CH3CN, 75% in two steps; iii)
Method (A): alkynes, Pd/C (2.5 mol%), CuI (15 mol%), DMF, O2, rt; iv) Method (B):
alkynes, NiCl2$6H2O, CuI (5 mol%), TMEDA (20 mol%), O2, THF, rt; v) MeOH, NH3, 16 h,
systems, we elaborated the synthesis of
a small library of
C5-substituted-1,3-diynyl-20-deoxyuridine by metal catalyzed
alkyne CeH activation on the C5-ethynyl intermediate 1 with
various alkynes (Scheme 1).
3
ꢀC.

222
O. Sari et al. / European Journal of Medicinal Chemistry 53 (2012) 220e228
reported as green concept. We tried to adjust and improve the
conditions to the cross-coupling reaction of phenylacetylene and
C5-ethynyl nucleoside 3 taken as a model; After optimization
(Table 1) of the phenylacetylene concentration (entry 2), solvent
(entry 3), temperature (entry 4) reaction time (entry 5) and
microwave activation (Biotage AB initiator EXP EU) (entry 6) the
reaction was carried out in the presence of Pd/C (2.5 mol%) and CuI
(15 mol%) in DMF under oxygen atmosphere and at rt (entry 1).
This procedure was successfully applied to other alkynes;
however, it failed or was less efficient with cyclo-alkynes and aryls
or alkynes bearing an amino or hydroxyl moiety; therefore, we
decided to realize the cross-coupling reaction, using the efficient
catalytic system for the heterocoupling of two terminal alkynes in
the presence of a nickel-copper co-catalyst recently reported by Yin
et al. [25]. The optimized conditions involving NiCl2$6H2O (5 mol%),
CuI (5 mol%) and TMEDA (20 mol%) in THF under oxygen atmo-
sphere have been selected to prepare a library of C5-substituted
1,3-diyne-20-deoxyuridines. The cross-coupling reaction between
the key intermediate 3 with different commercially available EDG-
(or EWG-)substituted aromatic and non-aromatic alkynes (Table 2)
gave in moderate to good yields (20e58%) fourteen nucleoside
analogues (4e17). It is noteworthy that under these conditions the
reaction occurred readily at room temperature after 16 h gave
better results and easier purification reaction mixture than opti-
mized palladium-mediated coupling reaction. As expected, the
efficiency of the coupling process depends directly on the alkyne
proton acidity. In fact, best yields were observed with EWD-
substituted aromatic alkynes (entries 8 and 9, Table 2) meanwhile
with ortho- (and para) EDG-substituted aromatic alkynes (entries 2,
3, 7, Table 2), yields are decreased. Otherwise, low yields were
observed in case of aliphatic-substituted alkynes (entries 11 and 13,
Table 2) probably due to the poor reactivity of the alkynes, except
for the entry 14 which gave surprisingly higher yield.
concentration required to reduce virus-induced cytopathicity by
50% (EC50). Data for varicella-zoster virus (VZV TKþ and TKꢁ),
human cytomegalovirus (AD-169 and Davis strains) in HEL cell
cultures are summarized in Table 3.
Among synthesized compounds, several of them showed
significant antiviral activities against several DNA viruses. Espe-
cially, with an EC50 of 2.8e3.7
mM for compound 19, 0.9e1.08 mM for
26 and 2.1e6.2 M for 29 against TK-positive (OKA) VZV strains,
m
these compounds were found similar or slightly more active than
acyclovir which was chosen as reference. This series of compounds
were found less active than brivudin against TKþ YS and OKA
strains, but compounds 19 and 20 are more efficient than the
references against the TKꢁ deficient (07/1) VZV strain, with EC50
values of 14 mM and 9.5 mM, respectively. For those data, it seems
that two groups differ by their effect, a tentative of SAR could be: (1)
For the group of nucleosides bearing aryl skeleton, a hydrophobic
tails (alkyl chain 19 (log P ¼ 2.36) and 20 (log P ¼ 3.70) or eOCF3 26
(log P ¼ 2.39)) appeared to be important for antiviral activity to
compare with the not active aryl (18) (log P ¼ 0.96), aminophenyl
(23, 24 (log P ¼ 0.13)) or nitro analogue (25) (log P ¼ 0.9); (2) Unlike
the first group, the substitution of the aryl by alkyl group shows
that the hydrophilic properties of the nucleoside for possible high
H-bonding (29) (log P ¼ ꢁ1.45) is suitable in this system than
cycloalkyl substituents (30 (log P ¼ 1.43), 31). Theses results
suppose that these two groups possess their own molecular
interactions in biological systems. The cytotoxic evaluation of the
most active compounds showed that they did not affect cell
morphology (MCC generally ꢂ100
m
M) and the CC50 values for
reduction of cell growth were usually ꢂ50
mM. However and
surprisingly, linear compounds 19 and 20 which were substituted
by a p-alkylphenyl group did not exhibit good anti-VZV replication
in contrast to the nM activities of their bicyclic furano pyrimidine
nucleoside derivatives [9]. We also evaluated the antiviral effect of
compounds 18e31 against a wide panel of other viral strains
including vaccinia virus (VV) and herpes simplex virus-1(KOS)
(HSV-1), HSV-1(TKꢁ), HSV-2(G) in HEL cell cultures, influenza
The deacetylation of 4e17 in methanolic ammonia solution
afforded the desired final compounds 18e31, respectively, which
were evaluated for their antiviral activities.
virus
A (H1N1 and H3N2) and B in MDCK cell cultures,
parainfluenza-3 virus, reovirus-1, Sindbis virus and Punta Toro
virus in Vero cell cultures and Vesicular stomatitis virus (VSV),
Coxsackie B4 and respiratory syncytial virus in HeLa cell cultures,
and HIV-1(IIIB) or HIV-2(ROD) in CEM cell cultures (data not
2.2. Antiviral activity
Compounds 18e31 were evaluated against a wide variety of
viruses, to determine their antiviral activity, as the effective
Table 1
Optimization of the Pd/C and CuI heterocoupling of nucleoside 3 with phenylacetylene.
O
O
NH
NH
AcO
N
O
AcO
O
N
Pd/C and CuI
(Table 1)
O
O
+
homodimer
OAc
OAc
3
4
Entry
Reaction time
Conditions
Yields for (4)
1
2
3
4
5
6
3 days
4 days
6 days
1 h 30
5 h
Pd/C, CuI, DMF, O2, rt, phenylacetylene (4 eq.)
Pd/C, CuI, DMF, O2, rt, phenylacetylene (8 eq.)
Pd/C, CuI, DMSO, O2, rt, phenylacetylene (4 eq.)
48% þ (3) þ homodimer
50% þ (3) þ homodimer
43% þ (3) þ homodimer
42% þ (3) þ homodimer þ degradation
Degradation
Pd/C, CuI, DMF, O2, 100 ꢀC, phenylacetylene (4 eq.)
Pd/C, CuI, DMF, O2, 100 ꢀC, phenylacetylene (4 eq.)
2 h
Pd/C, CuI, DMF, O2, 100 ꢀC, MW, phenylacetylene (4 eq.)
48% þ (3) þ homodimer

O. Sari et al. / European Journal of Medicinal Chemistry 53 (2012) 220e228
223
Table 2
Results for nickel-catalyzed hetero-coupling reaction of 3 with various alkynes.
R
R
O
O
N
O
N
NH
R
NH
NH
AcO
AcO
HO
NiCl2.6H2O (5 mol%)
MeOH/NH3
O
O
O
N
O
O
O
CuI (5 mol%)
TMEDA (20 mol%)
O2, THF, rt,16 h
OAc
OAc
4-17
OH
18-31
3
Entry
Re
Cross-coupling
Product number
Deacylated compounds
Isolated yield (%)
Product number
1
2
3
4
5
6
7
8
C6H5e
p-(C3H7)eC6H4e
p-(C6H13)eC6H4e
m-(MeO)2eC6H3e
2-Pyridinee
m-(NH2)C6H4e
p-(NH2)C6H4e
4
5
6
7
8
9
10
11
12
13
14
15
16
17
53
32
31
53
39
45
36
50
58
56
30
55
20
54
18
19
20
21
22
23
24
25
26
27
28
29
30
31
p-(NO2)C6H4e
9
p-(OCF3)C6H4e
m-(CH3)ep-(F)eC6H3e
CH3eCH(OH)eCH2e
HOCH2e
CyclopentaneeCH2e
Cyclopropyle
10
11
12
13
14
Table 3
Antiviral and cytotoxic activity of compounds 18e31 against, varicella-zoster virus (VZV TKþ and TKꢁ), human cytomegalovirus (AD-169 strain and Davis strain) in HEL cells.
MCCb
(m
M)
CC50
(mM)
a
a
c
Compound
EC50
(
m
M)
EC50
(m
M)
VZV (YS)
VZV (OKA)
TKꢁVZV (07/1)
CMV (AD-169)
CMV (Davis)
18
19
20
21
22
23
24
25
>20
2.77
6.44
38.07
2.06
e
>20
>20
>20
>20
>20
>100
49
>100
>100
>20
>20
>20
>20
>100
>4
>100
>100
>20
100
100
100
>100
>20
>100
>100
100
100
3.7 ꢃ 1.8
8.8 ꢃ 0.1
39 ꢃ 1.0
5.1 ꢃ 1.5
>100
14 ꢃ 0.3
9.5 ꢃ 0.2
>100
20
>100
>100
>20
52 ꢃ 9.1
35 ꢃ 6.1
>100
1.5 ꢃ 1.0
>100
e
>100
>20
>100
89
e
26
27
28
29
30
31
1.08
9.13
12.71
6.24
33.74
e
0.9 ꢃ 0.2
14 ꢃ 3.7
20 ꢃ 5.3
2.1 ꢃ 0.8
42 ꢃ 5.7
>100
>4
>20
>20
>20
>100
>100
>100
>100
e
>20
>20
>100
>100
>100
>100
e
100
100
55 ꢃ 1.1
45 ꢃ 11
9.5 ꢃ 6.6
>100
>100
>100
>100
>100
92 ꢃ 70
e
>100
>100
>100
>100
>300
>350
>150
>350
89 ꢃ 15
>100
Acyclovir
Ganciclovir
Brivudin
Cidofovir
2.18
e
2.5 ꢃ 0.3
470 ꢃ 490
130 ꢃ 86
>100
64 ꢃ 16
e
7.5 ꢃ 2.3
e
7.1 ꢃ 0
e
0.0075
e
0.0087
e
74
e
1.0 ꢃ 0.4
1.1 ꢃ 0
a
Effective concentration required to reduce virus-induced cytopathicity by 50%.
Minimum cytotoxic concentration that causes a microscopically detectable alteration of cell morphology.
Cytostatic concentration required to reduce cell growth by 50%.
b
c
shown). Only compound 26 exhibits moderate activity with EC50
implies that the compounds are able to penetrate into cells. Several
of the synthesized compounds showed activities against DNA
viruses in the nanomolar range, especially compound 5-[4-(4-
values against HSV-1: 14.5
14.5 M; and VV: 20 M.
mM; HSV-2: 10 m
M; HSV-1 (TKꢁ):
m
m
trifluoromethoxyphenyl)buta-1,3-diynyl]-20-deoxyuridine
(26)
which is the most potent inhibitor of this series against VZV with an
EC50 of w1 M and a CC50 of 55 M. This class of compounds could
3. Conclusions
m
m
Fourteen novel C5-(4-substituted-1,3-diyne) 20-deoxyuridines
were successfully synthesized through a nickel-copper catalyst.
They have been evaluated against several DNA viruses. Some
cellular activity was sometimes observed, which most likely
be of further interest for lead optimization as anti-infectious agents.
It will be also of interest to investigate the properties of their
monophosphorylated derivatives against thymidylate synthase and
their triphosphorylated derivatives against the viral and cellular

224
O. Sari et al. / European Journal of Medicinal Chemistry 53 (2012) 220e228
DNA polymerases to reveal the mechanism of biological (i.e. anti-
viral) action.
4.1.3. General procedure to compounds 4e17
To a THF solution of CuI (5 mol%) and NiCl2$6H2O (5 mol%),
tetramethylethylendiamine (20 mol%) was added. The solution was
stirred 2 min at room temperature. C5-Ethynyl-nucleoside (1 eq.)
and commercial alkyne (5 eq.) were added subsequently and the
reaction mixture was stirred overnight at room temperature under
oxygen atmosphere. After evaporation of all volatiles, the residue
was purified by silica gel column chromatography (EtOAc/Petro-
leum ether) to yield the desired compound.
4. Experimental section
4.1. Chemistry
Commercially available chemicals were of reagent grade and
used as received. The reactions were monitored by thin layer
chromatography (TLC) analysis using silica gel plates (Kieselgel
60F254, E. Merck). Column chromatography was performed on Silica
Gel 60 M (0.040e0.063 mm, E. Merck). The 1H and 13C NMR spectra
were recorded on a Varian InovaUnity 400 spectrometer (400 MHz)
in (d4) methanol, CDCl3, shift values in parts per million relative to
SiMe4 as internal reference. High Resolution Mass spectra were
performed by the Mass Spectrometry Centre of Blaise Pascal
University (Program Masslynx 4.0) (Aubière, France) and by the
“Fédération de Recherche” ICOA/CBM (FR2708) platform (Orléans,
France). Marvin, weighted, was used for calculate prediction of log
P, Marvin 5.3.4, 2010.
4.1.3.1. 30,50-Di-O-acetyl-5-(4-phenylbuta-1,3-diynyl)-20-deoxyur-
idine (4). Yield 53%, 1H NMR (400 MHz, CDCl3)
d 8.98 (s, 1H, NH),
0
7.98 (s, 1H, H6)0, 7.30e7.55 (m, 5H, HAr), 6.29 (t, 1H, J ¼ 6.0 Hz, H1 ),
0
0
0
5.23 (m, 1H, H3 ), 4.30e4.38 (m, 3H, H4 , 2H5 ), 2.60 (m, 2H, H2 ), 2.12
(s, 3H, OAc), 2.06 (s, 3H, OAc). 13C NMR (101 MHz, CDCl3)
d 170.2 (C]
O), 170.1 (C]O), 160.5 (C4), 148.7 (C2), 143.3 (C6), 132.5 (2ꢄ CAr0),
00
129.4 (CAr), 128.4 (2CAr), 121.2 (C4 ), 99.8 (C5), 85.7 (C4), 82.7 (C1 ),
30
50
20
78.6 (C00), 73.7 (C ), 73.3 (C00), 72.0 (C00), 63.6 (C ), 38.4 (C ), 20.9
(CH3, OAc), 20.8 (CH3, OAc). HRMS (ESI): m/z [M þ H]þ calcd for
C23H21N2O7: 437.134327, found: 437.134298. HRMS (ESI): m/z
[M þ Na]þ calcd for C23H20N2NaO7: 459.1163, found: 459.1160.
4.1.1. 30,50-Di-O-acetyl-5-iodo-20-deoxyuridine (2)
A solution of acetylated 20-deoxyuridine (1) (22 mmol) in dry
CH3CN (60 mL), CAN (7.21 g, 0.6 eq.) and I2 (3.33 g, 0.6 eq.) was
refluxed until completion (typically 1 h, checked by TLC). After
cooling to room temperature, solvents were evaporated under
reduced pressure and the dark oily residue was dissolved in AcOEt
(200 mL) and H2O (50 mL). The biphasic mixture was cooled in an
ice bath and a saturated Na2S2O3 solution was smoothly added until
complete decolouration. The organic layer was washed with water
(2 ꢄ 50 mL) and brine (50 mL), dried over MgSO4 and concentrated
under reduced pressure. The white foam was triturated with
pentane (50 mL), filtrated and dried under reduced pressure to
afford pure iodinated 2 in 87% yield. The physico-chemical data of 2
are in accordance to those previously published [24].
4.1.3.2. 30,50-Di-O-acetyl-5-[4-(4-propylphenyl)buta-1,3-diynyl]-20-
deoxyuridine (5). Yield 32%, 1H NMR (400 MHz, CDCl3)
d 8.91 (bs,
1H, NH), 7.98 (s, 1H, H6), 7.40 (d, J ¼ 8.1 Hz, 2H, HA0 r), 7.13 (d,
J ¼ 8.1 Hz, 2H, HAr), 6.29 (dd, J ¼ 7.5, 6.0 Hz, 1H, H1 ), 5.24 (dt,
0
0
J ¼ 6.6, 2.6 Hz, 1H, H3 ), 4.40 (dd, J ¼ 12.3, 3.0 Hz, 1H, H5a ), 4.34 (dd,
0
0
J ¼ 12.3, 2.9 Hz, 1H, H5b ), 4.30 (dd, J ¼ 5.7, 2.8 Hz, 1H, H4 ),
0
0
2.61e2.56 (m, 3H, H2a and CH2 alkyl), 2.29e2.19 (m, 2H, H2b ), 2.21
(s, 3H, OAc), 2.12 (s, 3H, OAc), 1.61 (h, J ¼ 7.6 Hz, 2H, CH2 alkyl),
0.92 (t, J ¼ 7.3 Hz, 3H, CH3 alkyl). 13C NMR (101 MHz, CDCl3)
d 170.5
(C]O), 170.3 (C]O), 160.8 (C4), 149.0 (C2), 144.8 (CAr), 143.4 (C60),
132.6 (2ꢄ CAr), 128.8 (2ꢄ CAr), 118.5 (CAr), 100.2 (C5), 85.9 (C1 ),
40
30
83.3 (C00), 82.9 (C ), 79.0 (C00), 73.9 (C ), 73.0 (C00), 71.9 (C00), 63.8
0
0
(C5 ), 38.6 (C2 ), 38.2 (CH2 alkyl), 24.4 (CH2 alkyl), 21.0 (CH3, OAc),
20.9 (CH3, OAc), 13.9 (CH3 alkyl). HRMS (ESI): m/z [M þ H]þ calcd
for C26H27N2O7: 479.1813, found: 479.1813.
4.1.2. 30,50-Di-O-acetyl-5-ethynyl-20-deoxyuridine (3)
Iodinated nucleoside 2 (3.4 mmol) was dissolved in a mixture of
dry DMF (15 mL), dry Et3N (1.44 mL, 3 eq.) and trimethylsilylace-
tylene (1.46 mL, 3 eq.). CuI (195 mg, 0.2 eq.) and PdCl2(PPh3)2
(240 mg, 0.1 eq.) were then added and the reaction mixture was
stirred at room temperature until completion (typically 5e20 h,
checked by TLC). Solvents were evaporated under reduced pres-
sure. The oily residue was dissolved in AcOEt (200 mL) then washed
with water (5 ꢄ 30 mL) and brine (40 mL). The organic layer was
dried over MgSO4 and the solvents were evaporated under reduced
pressure to dark oil. A purification using a short path flash chro-
matography (eluent: hexanes/AcOEt 7/3 then 1/1) afforded the
4.1.3.3. 30,50-Di-O-acetyl-5-[4-(4-hexylphenyl)buta-1,3-diynyl]-20-
deoxyuridine (6). Yield 31%, 1H NMR (400 MHz, CDCl3)
d 9.33 (s,
1H, NH), 7.98 (s, 1H, H6), 7.39 (d, J ¼ 8.1 Hz, 2H, HAr), 7.13 (d,
0
J ¼ 8.1 Hz, 2H, HAr), 6.32e6.27 (dd, J ¼ 7.3, 6.3, Hz, 1H, H1 ), 5.24
0
0
(dd, J ¼ 6.1, 2.8 Hz,1H, H3 ), 4.40 (dd, J ¼ 12.2, 3.0 Hz, 1H, H5a ), 4.33
0
0
(dd, J ¼ 12.4, 2.9 Hz, 1H, H5b ), 4.31e4.28 (m, 1H, H4 ), 2.64e2.53
0
0
(m, 3H, H2a , CH2eAr), 2.30e2.17 (m, 1H, H2b ), 2.20 (s, 3H, CH3,
OAc), 2.11 (s, 3H, CH3, OAc), 1.63e1.53 (m, 2H, CH2), 1.34e1.16 (m,
6H, 3ꢄ sCH2), 0.87 (t, J ¼ 6.7 Hz, 3H, CH3). 13C NMR (101 MHz,
CDCl3)
d
170.5 (C]O), 170.3 (C]O), 161.0 (C4), 149.1 (C2), 145.1
30,50-di-O-acetyl-5-trimethylsilylethynyl-20-deoxyuridine.
This
(CAr), 143.4 (C6), 132.6 (2ꢄ CAr), 128.7 (2ꢄ CAr), 118.5 (CAr), 100.1
10
40
30
compound (6.7 mmol), was dissolved in dry CH3CN (15 mL). TBAF
monohydrate (1.85 g, 1.05 eq.) was added and the resulting solution
was stirred at room temperature until completion (typically 30 min
to 2 h, checked by TLC). Solvents were evaporated under reduced
pressure at room temperature and the oily residue was submitted
to a flash column chromatography (eluent : hexanes/AcOEt 1/1
then AcOEt then MeOH/AcOEt 98/2) to afford pure 5-ethynyl
nucleoside 3 as white solid in 75% yield. mp ¼ 183 ꢀC. 1H NMR
(C5), 85.8 (C ), 83.2 (C00), 82.8 (C ), 78.9 (C00), 73.9 (C ), 73.0 (C00),
50
20
72.0 (C00), 63.8 (C ), 38.50 (C ), 36.1 (CH2 alkyl), 31.8 (CH2 alkyl),
31.2 (CH2 alkyl), 29.0 (CH2 alkyl), 22.7 (CH2 alkyl), 21.0 (CH3, OAc),
20.9 (CH3, OAc), 14.2 (CH3 alkyl). HRMS (ESI): m/z [M þ H]þ calcd
for C29H33N2O7: 521.2282, found: 521.2276.
4.1.3.4. 30,50-Di-O-acetyl-5-[4-(3,5-dimethoxyphenyl)buta-1,3-diynyl]-
20-deoxyuridine (7). Yield 53%, 1H NMR (400 MHz, CDCl3)
d 9.58 (s,
(400 MHz, CDCl3)
d
9.04 (s, 1H, NH), 7.93 (s, 1H, H6), 6.30 (t,
1H, NH), 7.98 (s, 1H, H6), 6.62 (d, J ¼ 2.3 Hz, 2H, HAr), 6.47 (t,
0
0
0
0
0
J ¼ 7.1 Hz, 1H, H1 ), 5.25 (m, 1H, H3 ), 4.30e4.45 (m, 3H, H40 and H5 ),
J ¼ 2.3 Hz, 1H, HAr), 6.27 (dd, J ¼ 7.4, 6.0 Hz, 1H, H1 ), 5.23 (dt, J ¼ 6.5,
0
0
0
3.21(s, 1H, CH alkyne), 2.55 (m, 1H, H2 a), 2.25 (m, 1H, H2 b), 2.07 (s,
2.4 Hz, 1H, H3 ), 4.39 (dd, J ¼ 12.2, 3.2 Hz, 1H, H5a ), 4.33 (dd, J ¼ 12.3,
0
0
3H, OAc), 2.05 (s, 3H, OAc); 13C NMR (101 MHz, CDCl3)
d
170.5,170.3,
3.0 Hz, 1H, H5b ), 4.29 (dd, J ¼ 5.7, 2.8 Hz, 1H, H4 ), 3.76 (s, J ¼ 2.9 Hz,
0
161.1, 149.2, 142.9, 99.8, 85.7, 82.4, 74.6, 74.0, 63.9, 38.5, 21.0, 20.9.
HRMS (ESI): m/z [M þ H]þ calcd for C15H17N2O7: 336.3052, found:
336.3048. The physico-chemical data of 3 are in accordance to
those previously published [24].
6H, 2ꢄ CH3eO), 2.57 (ddd, J ¼ 14.3, 5.8, 2.5 Hz, 1H, H2a ), 2.28e2.20
0
(m, 1H, H2b ), 2.19 (s, 3H, CH3, OAc), 2.10 (s, 3H, CH3, OAc). 13C NMR
(101 MHz, CDCl3)
d
170.5 (C]O), 170.4 (C]O), 161.2 (C4), 160.6 (2ꢄ
CAreO), 149.1 (C2), 143.7 (C6), 122.6 (CAr), 110.3 (2ꢄ CAr), 103.2 (CAr),

O. Sari et al. / European Journal of Medicinal Chemistry 53 (2012) 220e228
225
0
10
40
30
99.8 (C5), 85.92 (C ), 82.8 (C ), 82.7 (C00), 78.6 (C00), 73.9 (C ), 73.1
J ¼ 8.2 Hz, 2H, HAr), 6.27 (dd, J ¼ 7.3, 6.1 Hz, 1H, H1 ), 5.24 (dt, J ¼ 20.5
0
50
20
(C00), 72.5 (C00), 63.8 (C ), 55.6 (2ꢄ CH3eO), 38.5 (C ), 20.9 (2ꢄ CH3,
OAc). HRMS (ESI): HRMS (ESI): m/z [M þ H]þ calcd for C25H25N2O9:
497.1554, found: 497.1551.
and 5.8 Hz, 1H, H3 ), 4.36 (ddd, J ¼ 22.8,012.3 and 3.1 Hz, 1H, H5 ),
4.33e4.27 (m, J ¼ 5.8 and 2.9 Hz, 1H, H5 ), 2.58 (ddd,0 J ¼ 14.3, 5.8
0
and 2.5 Hz,1H, H2 ), 2.25 (dt, J ¼ 14.3 and 7.0 Hz,1H, H2 ), 2.18 (s, 3H,
CH3, OAc), 2.10 (s, 3H, CH3, OAc). 13C NMR (101 MHz, CDCl3)
d 170.5
4.1.3.5. 30,50-Di-O-acetyl-5-[4-(pyridin-2-yl)buta-1,3-diynyl]-20-deox-
(C]O), 170.3 (C]O), 161.2 (C4), 149.8 (CAr), 149.1 (C2), 143.9 (C6),
134.3 (2ꢄ CAr), 121.7 (CAr), 121.0 (2ꢄ CAr), 120.1 (OCF3), 99.6 (C5),
yuridine (8). Yield 39%, 1H NMR (400 MHz, CDCl3)
d 9.67 (s, 1H, NH),
0
00
30
8.57 (d, J ¼ 4.8 Hz, 1H, HAr), 8.01 (s, 1H, H6), 7.66 (d, J ¼ 8.5 Hz, 1H,
86.0 (C1 ), 82.9 (C4 ), 81.1 (C00), 78.3 (C00), 74.3 (C00), 73.9 (C ), 73.1
50
20
H
Ar), 7.64 (d, J ¼ 7.7 Hz, 1H, HAr), 7.26 (t, J ¼ 6.3 Hz, 1H, HAr), 6.28 (t,
(C00), 63.8 (C ), 38.5 (C ), 20.9 (CH3, OAc), 20.9 (CH3, OAc). HRMS
(ESI): m/z [M þ H]þ calcd for C24H20F3N2O8: 521.1166, found:
521.1161.
0
0
J ¼ 6.5 Hz, 1H, H1 ), 5.25e5.20 (m, 1H0, H3 ), 4.39 (dd, J ¼ 12.1, 02.9 Hz,
0
0
1H, H5a ), 4.34e4.29 (m, 2H, H5b , H4 ), 4.31e4.27 (m, 1H, H2 ), 2.25
0
(dt, 14.2, 7.1 Hz, 1H, H2 ), 2.18 (s, 3H, CH3, OAc), 2.09 (s, 3H, CH3, OAc).
13C NMR (101 MHz, CDCl3)
d
170.4 (C]O), 170.3 (C]O), 161.0 (C4),
4.1.3.10. 30,50-Di-O-acetyl-5-[4-(4-fluoro-3-methylphenyl)buta-1,3-
diynyl]-20-deoxyuridine (13). Yield 56%, 1H NMR (400 MHz, CDCl3)
150.4 (CAr), 149.1 (C2), 144.3 (C6), 141.9 (CAr), 136.3 (CAr), 132.0 (CAr),
10
40
123.8 (CAr), 99.4 (C5), 86.0 (C ), 82.9 (C ), 81.3 (C00), 78.1 (C00), 73.9
d
9.64 (s, 1H, NH), 7.92 (s, 1H, H6), 7.25 (t, J ¼ 7.6 Hz, 1H, HAr),
30
50
20
(C ), 73.6 (C), 73.3 (C00), 63.8 (C ), 38.5 (C ), 21.0 (CH3, OAc), 20.9
(CH3, OAc). HRMS (ESI): m/z [M þ H]þ calcd for C22H20N3O7:
438.1295, found: 438.1294.
7.24e7.20 (m,010 H, HAr), 6.88 (t, J ¼ 8.9 Hz, 1H, HAr), 6.22 (dd, J ¼ 7.2,
00
6.2 Hz, 1H, H1 ),0 5.18 (dt, J ¼ 6.6, 2.5 Hz, 1H, H3 ), 4.33 (dd, J ¼ 12.2,
0
3.2 Hz, 1H, H5a ), 4.27 (dd, J ¼ 12.3, 3.1 Hz, 1H, H5b ), 4.23 (dd,
0
0
J ¼ 5.7, 2.9 Hz, 1H, H40 ), 2.51 (ddd, J ¼ 14.3, 5.8, 2.5 Hz, 1H, H2a ),
2.25e2.15 (m, 1H, H2b ), 2.17 (d, J ¼ 1.6 Hz, 3H, CH3eAr), 2.13 (s, 3H,
4.1.3.6. 30,50-Di-O-acetyl-5-[4-(3-amino-phenyl)buta-1,3-diynyl]-20-
deoxyuridine (9). Yield 45%, 1H NMR (400 MHz, CDCl3)
d
7.98 (s, 1H,
CH3, OAc), 2.04 (s, 3H, CH3, OAc). 13C NMR (101 MHz, CDCl3)
d 170.5
H6), 7.08 (t, J ¼ 7.7 Hz, 1H, HAr), 6.87 (d, J ¼ 7.7 Hz, 1H, HAr), 6.78 (s,
(C]O), 170.4 (C]O), 162.0 (CAreF), 161.2 (C4), 149.2 (C2), 143.6
1H, HAr), 6.68 (dd, J ¼ 8.0 and 1.7 Hz, 1H, HAr0 ), 6.28 (dd, J ¼ 7.2,
(C6), 135.8 (CAr), 132.2 (CAr), 125.8 (CAr), 125.6 (CAr), 115.5 (CAr),
0
10
40
30
6.4 Hz, 1H, H1 ), 5.24 (dt, J ¼ 6.5, 2.4 Hz, 1H, H3 ),04.39 (dd, J ¼ 12.3,
99.8 (C5), 85.9 (C ), 82.8 (C ), 82.0 (C00), 78.6 (C00), 73.9 (C ), 72.9
0
50
20
3.1 Hz, 1H, H5a ), 4.33 (dd, J ¼ 12.3, 2.9 Hz, 1H, H5b ), 4.29 (dd, J ¼ 5.7,
(C00), 72.3 (C00), 63.8 (C ), 38.4 (C ), 20.9 (CH3, OAc), 20.9 (CH3,
OAc), 14.4 (CH3eAr). HRMS (ESI): m/z [M þ H]þ calcd for
C24H22FN2O7: 469.1405, found: 469.1401.
0
0
2.8 Hz, 1H, H4 ), 2.57 (ddd, J ¼ 14.2, 5.7 and 2.4 Hz, 1H, H2a ),
0
2.30e2.14 (m, 1H, H2b ), 2.19 (s, 3H, CH3), 2.10 (s, 3H, CH3). 13C NMR
(101 MHz, CDCl3):
d
170.5 (C]O),170.4 (C]O), 161.2 (C4),149.1 (C2),
146.5 (CAreNH2),143.6 (C6), 129.5 (CAr),123.0 (CAr), 121.9 (CAr),118.5
4.1.3.11. 30,50-Di-O-acetyl-5-(6-hydroxyhepta-1,3-diynyl)-20-deoxy-
10
40
(CAr), 116.7 (CAr), 100.0 (C5), 85.9 (C ), 83.3 (C00), 82.9 (C ), 78.8 (C00),
uridine (14). Yield 30%, 1H NMR (400 MHz, CDCl3)
d 9.45 (bs, 1H,
30
50
200
0
73.9 (C ), 72.8 (C00), 72.1 (C00), 63.8 (C ), 38.5 (C ), 21.0 (CH3, OAc),
20.9 (CH3, OAc). HRMS (ESI): m/z [M þ H]þ calcd for C23H22N3O7:
452.1452, found: 452.1450.
NH), 7.93 (s, 1H, H6), 6.26 (dd, J ¼ 7.4, 6.1 Hz, 1H, H1 ), 5.23 (dt,
0
0
J ¼ 6.6, 2.6 Hz, 1H, H3 ), 4.38 (dd, J ¼ 12.3, 3.3 Hz, 1H, H5a ), 4.32 (dd,
0
0
J ¼ 12.3, 3.0 Hz, 1H, H5b ), 4.28 (dd, J ¼ 5.9, 3.0 Hz, 1H, H4 ), 4.00 (h,
0
J ¼ 5.9 Hz, 1H, CH alkyl), 2.55 (ddd, J ¼ 14.3, 6.0, 2.5 Hz, 1H, H2a ),
4.1.3.7. 30,50-Di-O-acetyl-5-[4-(4-aminophenyl)buta-1,3-diynyl]-20-
2.50 (dd, J ¼ 5.7, 3.5 Hz, 2H, CH2 alkyl), 2.25 (dt, J ¼ 14.3, 7.1 Hz, 1H,
deoxyuridine (10). Yield 36%, 1H NMR (400 MHz, CDCl3)
d
7.95 (s,
H
2b), 2.17 (s, 3H, CH3, OAc), 2.10 (s, 3H, CH3, OAc), 1.28e1.23 (m, 3H,
1H, H6), 7.29 (d, J ¼ 8.4 Hz, 2H0, HAr), 6.58 (d, J ¼ 8.4 Hz, 2H, HAr),
CH3 alkyl). 13C NMR (101 MHz, CDCl3)
d 170.5 (C]O), 170.4 (C]O),
0 0
0
6.29 (dd, J ¼ 7.2, 6.2 Hz, 1H, H1 ), 5.26e5.22 (m, 1H, H3 ), 4.40 (dd,
161.3 (C4), 149.1 (C2), 143.7 (C6), 99.8 (C5), 85.9 (C1 ), 82.8 (C4 ), 82.6
0
0
30
J ¼ 12.3, 2.9 Hz, 1H, H5a ), 4.33 (dd, J ¼ 15.4, 3.0 Hz, 1H, H5b ), 4.30
(C00), 78.8 (C00), 74.0 (C ), 66.8 (C00), 66.3 (CeOH), 66.2 (C00), 63.8
0
0
0
(m, 1H, H4 ), 3.95 (bs, 2H, NH2), 2.56 (ddd, J ¼ 14.2, 5.7, 2.3 Hz, 1H,
(C5 ), 38.4 (C2 ), 30.2 (CH2 alkyl), 22.6 (CH3 alkyl), 21.0 (CH3, OAc),
20.9 (CH3, OAc). HRMS (ESI): m/z [M þ H]þ calcd for C20H23N2O8:
419.1448, found: 419.14485.
0
0
H
2a ), 2.28e2.15 (m, 1H, H2b ), 2.21 (s, 3H, CH3, OAc), 2.11 (s, 3H,
CH3, OAc). 13C NMR (101 MHz, CDCl3)
d 170.5 (C]O), 170.4 (C]O),
152.2 (CAr), 148.9 (C4), 148.0 (C2), 143.0 (C6), 134.3 (2ꢄ CAr), 114.7
10
40
(2ꢄ CAr), 110.2 (CAr), 100.1 (C5), 85.8 (C ), 84.2 (C00), 82.8 (C ), 79.5
4.1.3.12. 30,50-Di-O-acetyl-5-[5-hydroxypenta-1,3-diynyl]-20-deoxy-
30
50
20
(C00), 74.0 (C ), 71.7 (C00), 71.4 (C00), 63.9 (C ), 38.5 (C ), 21.0 (2ꢄ
CH3, OAc). HRMS (ESI): m/z [M þ H]þ calcd for C23H22N3O7:
452.1452, found: 452.1452.
uridine (15). Yield 55%, 1H NMR (400 MHz, CDCl3)
d
9.82 (s, 1H, NH),
0
0
7.98 (s, 1H, H6), 6.25 (t, J ¼ 6.6 Hz, 1H, H1 ), 5.206e5.22 (m, 1H, H3 ),
0
0
4.36 (m, 4H, H5a , H5b , CH2-O), 4.29 (m, 1H, H4 ),03.30 (bs, 1H, OH),
0
2.57 (dd, J ¼ 14.3, 3.8 Hz, 1H, H2 ), 2.31 (m, 1H, H2 ), 2.16 (s, 3H, CH3,
4.1.3.8. 30,50-Di-O-acetyl-5-[4-(4-nitrophenyl)buta-1,3-diynyl]-20-
OAc), 2.10 (s, 3H, CH3, OAc). 13C NMR (101 MHz, CDCl3)
d 170.7 (C]
0
deoxyuridine (11). Yield 50%, 1H NMR (400 MHz, CDCl3)
d
9.61 (s,
O), 170.6 (C]O), 161.6 (C4), 149.3 (C2), 144.5 (C6), 99.3 (C5), 86.0 (C1 ),
40
300
50
1H, NH), 8.19 (d, J ¼ 8.7 Hz, 1H, HAr), 8.02 0(s, 1H, H6), 7.63 (d,
82.8 (C ), 82.7 (C00), 77.9 (C00), 74.0 (C ), 69.6 (C00), 69.4 (C00), 63.9 (C ),
0
J ¼0 8.6 Hz, 1H, HAr), 6.26 (t, J ¼ 6.6 Hz, 1H, H1 ), 5.26e5.20 (m, 1H,
51.3 (CH2eOH), 38.3 (C2 ), 21.0 (2ꢄ CH3, OAc). HRMS (ESI): m/z
0
H3 ), 4.40 (dd, J ¼ 12.2, 3.2 Hz, 1H, H5 ), 4.35 (dd, J ¼ 12.5, 3.5 Hz,
[M þ H]þ calcd for C18H19N2O8: 391.1135, found: 391.1135.
0
0
1H, H5 ), 4.32e4.25 (m, 1H, H4 ), 2.59 (ddd, J ¼ 8.0, 6.9, 3.5 Hz, 1H,
0
0
H2 ), 2.25 (m, 1H, H2 ), 2.18 (s, 3H, CH3, OAc), 2.11 (s, 3H, CH3, OAc).
4.1.3.13. 30,50-Di-O-acetyl-5-[5-cyclopentylpenta-1,3-diynyl]-20-deox-
13C NMR (101 MHz, CDCl3)
d
170.5 (C]O), 170.29 (C]O), 161.0
yuridine (16). Yield 20%, 1H NMR (400 MHz, CDCl3) 0d 8.97 (bs, 1H,
(C4), 149.0 (C2), 147.7 (CAr), 144.3 (C6), 133.4 (2ꢄ CAr), 128.3 (CAr),
NH), 7.92 (s, 1H, H60), 6.28 (dd, J ¼ 7.4, 6.1 Hz, 1H, H1 ), 5.23 (dt, 1H,
10
40
0
123.8 (2ꢄ CAr), 99.2 (C5), 86.1 (C ), 82.9 (C ), 80.3 (C00), 78.3 (C00),
J ¼ 6.0, 2.6 Hz, H3 ), 4.39 (dd, J ¼ 12.3, 3.0 Hz, 1H, H50 ), 4.32 (dd,
30
50
20
0
77.8 (C00), 75.4 (C00), 73.8 (C ), 63.8 (C ), 38.6 (C ), 20.9 (2ꢄ CH3,
OAc). HRMS (ESI): m/z [M þ H]þ calcd for C23H20N3O9: 482.1194,
found: 482.1188.
J ¼ 12.3, 2.9 Hz, 1H, H5 ), 4.29 (dd, J ¼ 5.7, 2.9 Hz, 1H, H4 ), 2.54 (ddd,
0
J ¼ 14.3, 5.9, 2.6 Hz, 1H, H2 ), 2.33 (d, J ¼ 6.8 Hz, 2H, CH2 alkyl),
0
2.26e2.15 (m, 1H, H2 ), 2.18 (s, 3H, CH3, OAc), 2.11 (s, 3H, CH3, OAc),
2.13e2.02 (m, 1H, CH alkyl), 1.84e1.74 (m, 2H, CH2 alkyl), 1.68e1.57
(m, 2H, CH2 alkyl), 1.57e1.49 (m, 2H, CH2 alkyl), 1.27e1.22 (m, 2H,
4.1.3.9. 30,50-Di-O-acetyl-5-[4-(4-trifluoromethoxyphenyl)buta-1,3-
diynyl]-20-deoxyuridine (12). Yield 58%, 1H NMR (400 MHz, CDCl3)
CH2 alkyl). 13C NMR (101 MHz, CDCl3)
d
170.5 (C]O, OAc), 170.3 (C]
d
9.68 (s,1H, NH), 8.00 (s,1H, H6), 7.51 (d, J ¼ 8.8 Hz, 2H, HAr), 7.16 (d,
O, OAc), 161.1 (C4), 149.0 (C2), 143.3 (C6), 100.3 (C5), 85.9 (C00), 85.7

226
10
O. Sari et al. / European Journal of Medicinal Chemistry 53 (2012) 220e228
40
30
50
(C ), 82.8 (C ), 79.4 (C000), 73.9 (C ), 65.2 (C00), 64.8 (C00), 63.9 (C ),
38.8 (CH alkyl), 38.5 (C2 ), 32.2 (2ꢄ CH2 alkyl), 25.4 (CH2 alkyl), 25.3
(2ꢄ CH2 alkyl), 21.0 (CH3, OAc), 20.9 (CH3, OAc). HRMS (ESI): m/z
[M þ H]þ calcd for C23H27N2O7: 443.1812, found: 443.1810.
4.1.4.4. 5-[4-(3,5-Dimethoxyphenyl)buta-1,3-diynyl]-20-deoxyur-
idine (21). Yield 73%, 1H NMR (400 MHz, MeOD) 8.52 (s, 1H, H6),
d
6.64 (d, J ¼ 2.3 Hz,0 2H, HAr), 6.54 (t, J ¼ 2.3 Hz, 1H, HA0 r), 6.23 (t,
J ¼ 6.4 Hz, 1H, H1 ), 4.41 (dt, J ¼ 6.0, 3.7 Hz, 1H, H3 ), 3.96 (q,
0
0
J ¼ 3.2 Hz, 1H, H4 ), 3.85 (dd, J ¼ 12.0, 2.9 Hz, 01H, H5a ), 3.77 (s, 6H,
4.1.3.14. 30,50-Di-O-acetyl-5-[4-(cyclopropyl)buta-1,3-diynyl]-20-
2ꢄ CH3eO), 3.75 (dd, J ¼ 12.0, 3.4 Hz, 1H, H5b ), 2.34 (ddd, J ¼ 13.6,
0
0
deoxyuridine (17). Yield 54%, 1H NMR (400 MHz, CDCl3)
d
9.03 (s,
6.1, 3.9 Hz, 1H, H2a ), 2.30e2.20 (m, 1H, H2b ). 13C NMR (101 MHz,
0
1H, NH), 7.90 (s, 1H, H6), 6.27 (dd, J ¼ 7.6, 5.9 Hz, 1H, H10 ), 5.22 (dt,
MeOD)
d
164.2 (C4), 162.3 (2ꢄ CAreO), 151.0 (C2), 147.4 (C6), 1240.0
0
0
0
J ¼ 6.6, 2.6 Hz, 1H, H3 ), 4.38 (dd, J ¼ 12.2, 3.0 Hz, 1, H5a ), 4.32 (dd,
(CAr), 111.1 (2ꢄ CAr), 103.7 (CAr), 99.1 (C5 ), 89.2 (C1 ), 87.4 (C4 ),
0
0
30
50
J ¼ 12.3, 3.0 Hz, 1H, H5b ), 4.30 (dd, J ¼ 5.6, 2.8 Hz, 1H, H4 ), 2.54
82.6 (C00), 78.0 (C00), 74.5 (C00), 73.9 (C00), 71.8 (C ), 62.4 (C ), 56.0
0
0
0
(ddd, J ¼ 14.3, 5.8, 2.6 Hz, 1H, H2a ), 2.25e2.18 (m, 1H, H2b ), 2.17
(s, 3H, CH3), 2.11 (s, 3H, CH3), 1.36 (m, 1H, CH alkyl), 0.95e0.76
(2ꢄ CH3eO), 41.9 (C2 ). HRMS (ESI): m/z [M þ Na]þ calcd for
C21H20N2O7Na: 435.1168, found: 435.1185.
(m, 4H, 2ꢄ CH2). 13C NMR (101 MHz, CDCl3)
d 170.5 (C]O),
170.3 (C]O), 161.1 (C4), 149.0 (C2), 143.2 (C6), 100.3 (C5), 89.4
4.1.4.5. 5-[4-(Pyridin-2-yl)buta-1,3-diynyl]-20-deoxyuridine
(22).
d
Yield 73%, 1H NMR (400 MHz, MeOD) 8.60 (s, 1H, H6), 8.57e8.51
10
40
30
50
(C00), 85.7 (C ), 82.8 (C ), 79.6 (C00), 74.0 (C ), 64.8 (C00), 63.9 (C ),
20
60.1 (C00), 38.5 (C ), 21.0 (CH3, OAc), 20.9 (CH3, OAc), 9.3 (CH2
(m, 1H, HAr), 7.86 (td, J ¼ 7.8, 1.7 Hz, 1H, HAr), 7.63 (d, J ¼ 7.9 Hz,
alkyl), 0.4 (CH alkyl). HRMS (ESI): m/z [M þ H]þ calcd for
1H, HAr), 7.45 (ddd, J ¼ 7.7, 5.0,1.0 Hz,1H, HAr), 6.23 (t, J ¼ 6.4 Hz,1H,
0
0
0
C20H21N2O7: 401.1343, found: 401.1339.
H1 ), 4.42 (dt, J ¼ 6.0, 3.7 Hz, 1H, H3 ), 3.96 (q, J ¼ 3.2 Hz, 1H, H4 ),
0
3.85 (dd, J ¼ 12.0, 2.9 Hz, 1H, H5a ), 3.75 (dd, J ¼ 12.0, 3.3 Hz, 1H,
0
0
0
4.1.4. General procedure to 18e31
H
5b ), 2.35 (ddd, J ¼ 13.6, 6.1, 4.0 Hz, 1H, H2 ), 2.30e2.21 (m, 1H, H2 ).
Compounds 4e17 were respectively dissolved in a solution of
NH3 in methanol (7 N). The mixture was stirred at 3 ꢀC overnight.
After evaporation of the solvents, the residual solid was purified on
silica gel eluting with CH2Cl2/MeOH (98:2) to give compounds
18e31, respectively.
13C NMR (101 MHz, MeOD) 164.1 (C4), 151.3 (C2), 151.0 (CAr), 148.2
d
(C6), 143.0 (CAr), 138.6 (CAr), 129.7 (CAr), 125.5 (CAr), 104.1, 98.5 (C5),
10
40
89.3 (C ), 87.5 (C ), 80.7 (C"), 77.2 (C00), 76.3 (C00), 74.8 (C00), 71.8
0
0
0
(C3 ), 62.4 (C5 ), 42.0 (C2 ). HRMS (ESI): m/z [M þ H]þ calcd for
C18H16N3O5: 354.1090, found: 354.1087.
4.1.4.1. 5-(4-Phenylbuta-1,3-diynyl)-20-deoxyuridine (18). Yield 68%,
4.1.4.6. 5-[4-(3-Amino-phenyl)buta-1,3-diynyl]-20-deoxyuridine (23).
1H NMR (400 MHz, MeOD)
d
8.53 (s, 1H, H6), 7.53e7.46 (m, 2H, HAr),
Yield 65%, 1H NMR (400 MHz, MeOD) 8.51 (s, 1H, H6), 7.07 (t,
d
0
7.44e7.32 (m, 3H, HAr), 6.24 (t, J ¼ 6.4 Hz, 1H, H1 ), 4.41 (dt, J ¼ 6.0,
J ¼ 7.8 Hz, 1H, HAr), 6.81 (s, 1H, HAr), 6.79 (dd, J ¼ 7.6, 0.9 Hz, 1H, HAr),
0
0
0
3.7 Hz,1H, H3 ), 3.96 (q, J ¼ 3.2 Hz,1H, H4 ), 3.85 (dd, J ¼ 12.0, 2.9 Hz,
6.76e6.72 (m, 1H, HAr), 6.23 (t, J ¼ 6.4 Hz, 1H, H1 ), 4.41 (dt, J ¼ 6.0,
0
0
0
0
1H, H5a ), 3.75 0(dd, J ¼ 12.0, 3.2 Hz, 1H,0H5b ), 2.34 (ddd, J ¼ 12.0, 6.4,
3.2 Hz,1H, H2a ), 2.30e2.21 (m,1H, H2b ). 13C NMR (101 MHz, MeOD)
3.6 Hz, 1H, H3 ), 3.95 (q, J ¼ 3.2 Hz, 1H, H4 ),03.84 (dd, J ¼ 12.0, 3.8 Hz,
0
1H, H5a ), 3.75 0(dd, J ¼ 12.0, 3.3 Hz, 1H,0 H5b ), 2.34 (ddd, J ¼ 13.5, 6.1,
d
164.1 (C4), 151.0 (C2), 147.4 (C6), 133.4 (2ꢄ CAr), 130.6 (CAr), 129.7
3.9 Hz, 1H, H2a ), 2.29e2.21 (m, 1H, H2b ). 13C NMR (101 MHz, MeOD)
10
40
(2ꢄ CAr), 122.8 (CAr), 99.1 (C5), 89.3 (C ), 87.3 (C ), 82.5 (C00), 78.0
d
: 164.2 (C4), 151.0 (C2), 149.3 (CAr, CeNH2), 147.2 (C6), 130.3 (CAr0),
30
50
20
(C00), 74.5 (C00), 74.4 (C00), 71.8 (C ), 62.4 (C ), 41.9 (C ). HRMS (ESI):
123.0 (CAr), 122.8 (CAr), 119.3 (CAr), 117.7 (CAr), 99.3 (C5), 89.2 (C4 ),
10
30
50
m/z [M þ Li]þ calcd for C19H16LiN2O5: 359.12142, found: 359.12190.
87.3 (C ), 83.5 (C00), 78.3 (C00), 73.9 (C00), 73.3 (C00), 71.8 (C ), 62.4 (C ),
0
41.9 (C2 ). HRMS (ESI): m/z [M þ H]þ calcd for C19H18N305: 368.1246,
4.1.4.2. 5-[4-(4-Propylphenyl)buta-1,3-diynyl]-200-deoxyuridine (19).
found: 368.1232.
Yield 66%, 1H NMR (400 MHz, MeOD)
d
8.52 (s, 1H, H6), 7.40 (d,
J ¼0 8.2 Hz, 2H, HAr), 7.19 (d, J ¼ 8.2 Hz, 2H, HAr), 6.24 (t, J ¼ 6.4 Hz, 1H,
4.1.4.7. 5-[4-(4-Aminophenyl)buta-1,3-diynyl]-20-deoxyuridine (24).
0
0
H1 ), 4.41 (dt, J ¼ 6.0, 3.6 Hz,1H, H3 ), 3.95 (q, J ¼ 3.2 Hz, 1H, H4 ), 3.84
Yield 72%, 1H NMR (400 MHz, MeOD) 8.45 (s, 1H, H6), 7.21 (d,
d
0
0
(dd, J ¼ 12.0, 2.9 Hz, 1H, H5a ), 3.75 (dd, J ¼ 12.0, 3.3 Hz, 1H, H5b ), 2.61
J ¼0 8.7 Hz, 2H, HAr), 6.61 (d, J ¼ 8.7 Hz, 2H, HAr), 6.24 (t, J ¼ 6.5 Hz,1H,
0
(t, J ¼ 7.4 Hz, 2H, CH2 alkyl), 2.34 (ddd, J ¼ 13.6, 6.2, 3.9 Hz, 1H, H2a ),
0
0
0
H1 ), 4.41 (dt, J ¼ 6.0, 3.6 Hz, 1H, H3 ), 3.95 (q, J ¼ 3.2 Hz, 1H, H4 ), 3.84
0
0
2.29e2.20 (m, 1H, H2b ), 1.64 (h, J ¼ 7.6 Hz, 2H, CH2 alkyl), 0.93 (t,
(dd, J ¼ 12.0, 2.9 Hz, 1H, H5a ), 3.75 (dd, J ¼ 12.0, 3.4 Hz, 1H, H5b ), 2.33
0
0
J ¼ 7.4 Hz, 3H, CH3 alkyl). 13C NMR (101 MHz, MeOD)
d
164.3 (C4),
(ddd, J ¼ 13.5, 6.1, 3.9 Hz, 1H, H2a ), 2.29e2.20 (m, 1H, H2b ). 13C NMR
151.1 (C2), 147.4 (C6), 146.1 (2ꢄ CAr), 133.5 (2ꢄ CAr), 130.0 (CAr), 120.1
(101 MHz, MeOD)
d
164.4 (C4), 151.3 (C2), 151.2 (CAr), 146.7 (C6), 1350.0
40
50
10
20
0
(CAr), 99.4 (C5), 89.4 (C ), 87.5 (C ), 83.2 (C00), 78.4 (C00), 74.3 (C00), 74.1
(2ꢄ CAr), 115.5 (2ꢄ CAr), 109.9 (CAr), 100.0 (C5), 89.3 (C4 ), 87.4 (C1 ),
30
30
50
(C00), 72.0 (C ), 62.6 (C ), 42.1 (C ), 39.1 (CH2 alkyl), 25.6 (CH2 alkyl),
14.1 (CH3 alkyl). HRMS (ESI): m/z [M þ Na]þ calcd for C22H22N2O5Na:
417.1426, found: 417.1430.
84.7 (C00), 79.2 (C00), 73.2 (C00), 72.34 (C00), 72.1 (C ), 62.6 (C ), 42.0
0
(C2 ). HRMS (ESI): m/z [M þ Na]þ calcd for C19H17N3O5Na: 390.1066,
found: 390.1077.
4.1.4.3. 5-[4-(4-Hexylphenyl)buta-1,3-diynyl]-200-deoxyuridine (20).
4.1.4.8. 5-[4-(4-Nitrophenyl)buta-1,3-diynyl]-20-deoxyuridine (25).
Yield 71%, 1H NMR (400 MHz, MeOD)
d
8.52 (s, 1H, H6), 7.40 (d, 2H,
Yield 54%, 1H NMR (400 MHz, MeOD) 8.60 (s, 1H, H6), 8.25 (d,
d
J ¼ 8.2 Hz, HAr), 7.19 (d, 2H, J ¼ 8.2 Hz, HAr), 6.24 (t, 1H, J ¼ 6.4 Hz,
J ¼ 8.9 Hz, 2H, HAr), 7.73 (d, J ¼ 8.9 Hz, 2H, HAr), 6.23 (t, J ¼ 6.4 Hz,
0
0
0
0
0
0
H1 ), 4.41 (dt, 1H, J ¼ 6.2 and 3.7 Hz, H3 ), 3.95 (q, 1H,J ¼ 3.2 Hz, H4 ),
1H, H1 ), 4.41 (dt, J ¼ 6.0, 3.8 Hz, 1H, H3 ), 3.96 (q, J ¼ 3.2 Hz, 1H, H4 ),
0
0
3.84 (dd, 1H, J ¼ 12.0 and 2.9 Hz, H5a ), 3.75 (dd, 1H, J ¼ 12.0, 3.3 Hz,
3.85 (dd, J ¼ 12.0, 2.9 Hz, 1H, H5 a), 3.75 (dd, J ¼ 12.2, 3.4 Hz, 1H,
0
0
0
H
5b ), 2.63 (t, 2H, J ¼ 7.5 Hz, CH2eAr), 2.34 (ddd, 1H, J ¼ 13.6, 6.1 and
H
H
5 b), 2.36 (ddd, J ¼ 13.6, 6.2, 4.0 Hz, 1H, H2 a), 2.31e2.21 (m, 1H,
0
0
0
3.9 Hz, H2a ), 2.25 (m,1H, H2b ),1.64e1.57 (m, 2H, CH2 alkyl),1.31 (m,
8H, 4ꢄ CH2 alkyl), 0.89 (t, 3H, J ¼ 6.8 Hz, CH3 alkyl). 13C NMR
2 b). 13C NMR (101 MHz, MeOD)
d
164.2 (C4), 151.2 (C2), 149.4 (CAr),
148.4 (C6), 134.8 (2ꢄ CAr), 129.9 (CAr), 125.1 (2ꢄ CAr), 98.7 (C5), 89.5
40
10
30
(101 MHz, MeOD)
d
164.2 (C4), 151.0 (C2), 147.2 (C6), 146.2 (CAr),
(C ), 87.6 (C ), 86.4 0(C00), 80.5 (C00), 79.1 (C00), 77.5 (C00), 72.2 (C ),
0
0
133.4 (2ꢄ CAr), 129.8 (2ꢄ CAr), 119.9 (CAr), 99.3 (C5), 89.2 (C4 ), 87.3
62.5 (C5 ), 42.2 (C2 ). HRMS (ESI): m/z [M þ Na]þ calcd for
10
30
50
(C ), 82.9 (C00), 78.2 (C00), 74.1 (C00), 73.9 (C00), 71.9 (C ), 62.4 (C ),
C19H15N3O7Na: 420.0808, found: 420.0820.
0
41.9 (C2 ), 36.9 (CH2 alkyl), 32.8 (CH2 alkyl), 32.4 (CH2 alkyl), 30.0
(CH2 alkyl), 23.6 (CH2 alkyl), 14.4 (CH3 alkyl). HRMS (ESI): m/z
4.1.4.9. 5-[4-(4-Trifluoromethoxyphenyl)buta-1,3-diynyl]-20-deoxy-
[M þ Na]þ calcd for C25H28N2O5Na: 459.1896, found: 459.1907.
uridine (26). Yield 71%, 1H NMR (400 MHz, MeOD) 8.55 (s,1H, H6),
d

O. Sari et al. / European Journal of Medicinal Chemistry 53 (2012) 220e228
227
0
7.61 (d, J ¼ 8.9 Hz,0 2H, HAr), 7.30 (d, J ¼ 8.1 Hz, 2H, HA0 r), 6.23 (t,
H2b ), 0.93e0.84 (m, 3H, CH and CH2 alkyl), 0.77e0.71 (m, 2H, CH2
J ¼ 6.4 Hz, 1H, H1 ), 4.41 (dt, J ¼ 6.0, 3.8 Hz, 1H, H30 ), 3.96 (q,
alkyl). 13C NMR (101 MHz, MeOD)
d
164.4 (C4), 151.0 (C2), 147.0 (C6),
0
40
10
30
J ¼ 3.2 Hz, 1H, H4 ), 3.85 (dd, J ¼ 12.0, 2.9 Hz, 1H, H5a ), 3.75 (dd,
99.5 (C5), 89.2 (C ), 89.0 (C00), 87.20 (C ), 79.1 (C00), 71.9 (C ), 66.6
0
0
50
300
J ¼ 12.0, 3.3 Hz, 1H, H5b ), 2.35 (ddd, J ¼ 13.6, 6.1, 3.9 Hz, 1H, H2a ),
(C00), 62.4 (C ), 61.06 (C ), 41.84 (C2 ), 9.43 (CH2 and CH, alkyl), 0.75
(CH2, alkyl). HRMS (ESI): m/z [M þ Na]þ calcd for C16H16N205Na:
339.0957, found: 339.0941.
0
2.30e2.21 (m, 1H, H2b ). 13C NMR (101 MHz, MeOD)
d
164.1 (C4),
151.17 (CAreO) 150.9 (C2), 147.7 (C6), 135.3 (2ꢄ CAr), 122.5 (2ꢄ CAr),
10
40
122.2 (OCF3, CAr), 98.9 (C5), 89.3 (C ), 87.4 (C ), 80.9 (C00), 77.7 (C00),
30
50
20
75.4 (C00), 75.2 (C00), 71.8 (C ), 62.4 (C ), 42.0 (C ). HRMS (ESI): m/z
[M þ Na]þ calcd for C20H15N2O6F3Na: 459.0780, found: 459.0775.
4.2. Antiviral activity assays
The antiviral assays, other than the anti-HIV assays, were based
on inhibition of virus-induced cytopathicity or plaque formation in
HEL [herpes simplex virus type 1 (HSV-1) (KOS), HSV-2 (G), vaccinia
virus, vesicular stomatitis virus, cytomegalovirus (HCMV), and
varicella-zoster virus (VZV)], Vero (parainfluenza-3, reovirus-1,
Sindbis virus and Coxsackie B4), HeLa (vesicular stomatitis virus,
Coxsackie virus B4, and respiratory syncytial virus) or MDCK
[influenza A (H1N1; H3N2) and influenza B] cell cultures. Confluent
cell cultures (or nearly confluent for MDCK cells) in microtiter 96-
well plates were inoculated with 100 CCID50 of virus (1 CCID50
being the virus dose to infect 50% of the cell cultures) or with 20
plaque forming units (PFU). After a 1e2 h virus adsorption period,
residual virus was removed, and the cell cultures were incubated in
4.1.4.10. 5-[4-(4-Fluoro-3-methylphenyl)buta-1,3-diynyl]-20-deoxy-
uridine (27). Yield 62%, 1H NMR (400 MHz, MeOD) 8.52 (s,1H, H6),
d
7.40 (d, J ¼ 7.3 Hz,1H, HAr), 7.37e7.31 (m,1H, HAr), 7.08e7.00 (m,1H,
0
0
H
Ar), 6.23 (t, J ¼ 6.4 Hz,1H, H1 ), 4.41 (dt, J ¼ 6.0, 3.7 Hz,1H, H3 ), 3.95
0
0
(q, J ¼ 3.2 Hz, 1H, H4 ), 30.84 (dd, J ¼ 12.0, 2.9 Hz, 1H, H5a ), 3.75 (dd,
0
J ¼ 12.0, 3.4 Hz, 1H, H50 b ), 2.34 (ddd, J ¼ 13.6, 6.1, 3.9 Hz, 1H, H2a ),
2.29e2.21 (m, 1H, H2b ), 2.25 (s, 3H, AreCH3). 13C NMR (101 MHz,
MeOD)
d
164.1 (C4), 151.0 (C2), 147.4 (C6), 136.7 (CAr), 133.2 (CAr0),
0
127.0 (CAr), 126.8 (CAr), 118.8 (CAr), 116.6 (CAr), 99.1 (C5 ), 89.3 (C1 ),
40
30
87.3 (C ), 81.78 (C00), 78.02 (C00), 74.33 (C00), 73.9 (C00), 71.8 (C ), 62.4
0
0
(C5 ), 41.9 (C2 ), 22.0 (CH3). HRMS (ESI): m/z [M þ Na]þ calcd for
C20H17N2O5FNa: 407.1019, found: 407.1028.
the presence of varying concentrations (200, 40, mM) of the test
4.1.4.11. 5-(6-Hydroxyhepta-1,3-diynyl)-20-deoxyuridine (28). Yield
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. Antiviral activity was expressed
as the EC50 or compound concentration required to reduce virus-
induced cytopathogenicity or viral plaque (VZV) plaque formation
by 50%. The minimal cytotoxic concentration (MCC) of the
compounds was defined as the compound concentration that
caused a microscopically visible alteration of cell morphology.
Alternatively, the cytostatic activity of the test compounds was
measured based on inhibition of cell growth. HEL cells were seeded
at a rate of 5 ꢄ 103 cells/well into 96-well microtiter plates and
allowed to proliferate for 24 h. Then, medium containing different
concentrations of the test compounds was added. After 3 days of
incubation at 37 ꢀC, the cell number was determined with a Coulter
counter. The cytostatic concentration was calculated as the CC50, or
the compound concentration required to reduce cell proliferation
by 50% relative to the number of cells in the untreated controls. The
methodology of the anti-HIV assays was as follows: human CEM
(w3 ꢄ 105 cells/cm3) cells were infected with 100 CCID50 of
65%, 1H NMR (400 MHz, MeOD)
d
8.43 0(s, 1H, H6), 6.22 (t, J ¼ 6.4 Hz,
0
0
1H, H1 ), 4.40 (dt, J ¼ 6.0, 3.4 Hz,01H, H3 ), 3.94 (q, J ¼ 3.2 Hz, 1H, H4 ),
0
3.82 (dd, J ¼ 12.0, 3.0 Hz, 1H, H5 ), 3.73 (dd, J ¼ 12.0, 3.4 Hz, 1H, H5 ),
2.49 (dd, J ¼ 8.1, 5.9 Hz, 2H, CH2 alkyl), 2.32 (ddd, J ¼ 13.6, 6.1,
0
0
3.9 Hz, 1H, H2 ), 2.28e2.18 (m, 1H, H2 ), 1.98 (bs, 1H, CH alkyl), 1.25
(d, J ¼ 6.2 Hz, 3H, CH3 alkyl). 13C NMR (101 MHz, MeOD)
d
164.3 (C4),
40
10
151.0 (C2), 147.1 (C6), 99.4 (C5), 89.2 (C ), 87.2 (C ), 82.7 (C00), 78.7
30
50
(C00), 71.9 (C ), 67.7 (C00), 67.3 (C00), 67.1 (CH alkyl), 62.4 (C ),
0
41.8(C2 ), 30.3 (CH2 alkyl), 22.6 (CH3 alkyl). HRMS (ESI): m/z
[M þ Na]þ calcd for C16H18N206Na: 357.1063, found: 357.1058.
4.1.4.12. 5-[5-Hydroxypenta-1,3-diynyl]-20-deoxyuridine (29). Yield
52%, 1H NMR (400 MHz, MeOD)
d
8.48 (s, 1H, H6), 6.22 (t, J ¼ 6.4 Hz,
0
0
1H, H1 ), 4.40 (dt, J ¼ 6.0, 3.7 Hz, 1H, H3 ), 4.29 (s, 2H, CH2eO), 3.94
0
0
(q, J ¼ 3.2 Hz, 1H, H4 ), 3.83 (dd, J ¼ 12.0, 3.0 Hz, 1H, H5a ), 3.74(dd,
0
0
J ¼ 12.0, 3.4 Hz, 1H, H50b ), 2.33 (ddd, J ¼ 13.6, 6.2, 3.9 Hz, 1H, H2a ),
2.28e2.19 (m, 1H, H2b ). 13C NMR (101 MHz, MeOD)
d
164.2 (C4),
50
10
40
151.0 (C2), 147.5 (C6), 99.0 (C ), 89.2 (C ), 87.3 (C ), 83.0 (C00), 77.8
30
20
(C00), 71.8 (C ), 70.8 (C00), 69.7 (C00), 62.4 (C5), 51.1 (CH2), 41.9 (C ).
HRMS (ESI): m/z [M þ Na]þ calcd for C14H14N2O6Na: 329.0750,
found: 329.0739.
HIV(IIIB) or HIV-2(ROD)/mL and seeded in 200
mL wells of
a microtiter plate containing appropriate dilutions of the test
compounds. After 4 days of incubation at 37 ꢀC, HIV-induced CEM
giant cell formation was examined microscopically.
4.1.4.13. 5-[5-Cyclopentylpenta-1,3-diynyl]-20-deoxyuridine
(30).
Yield 62%, 1H NMR0 (400 MHz, MeOD)
d
8.42 (s, 1H, H6), 6.22 (t,
Acknowledgements
0
J ¼ 6.5 Hz, 1H, H1 ), 4.40 (dt, J ¼ 6.0 3.6 Hz, 1H, H30 ), 3.94 (q,
0
J ¼ 3.2 Hz, 1H, H4 ), 3.83 (dd, J ¼ 12.0, 3.0 Hz, 1H, H5a ), 3.73 (dd,
O.S. is grateful to the French Ministère de l’Enseignement
Supérieur MNERST for a Ph.D. scholarship. The research was sup-
ported by the KU Leuven (GOA 10/014). The authors thank L. Per-
soons, F. De Meyer, L. Ingels, A. Camps, L. Van den Heurck and S.
Carmans for dedicated technical assistance for the antiviral assays.
0
J ¼ 12.0, 3.4 Hz, 1H, 0 H5b ), 2.36 (d, J ¼ 6.7 Hz, 2H, CH2 alkyl),
0
2.34e2.28 (m, 1H, H2a ), 2.27e2.18 (m, 1H, H2b ), 2.13e2.02 (m, 1H,
CH alkyl), 1.85e1.78 (m, 2H, CH2 alkyl), 1.71e1.53 (m, 4H, 2ꢄ CH2
alkyl), 1.34e1.26 (m, 2H, CH2 alkyl). 13C NMR (101 MHz, MeOD)
0
0
d
164.4 (C4), 151.0 (C2), 146.9 (C6), 99.5 (C5), 89.2 (C4 ), 87.2 (C1 ), 85.4
30
50
20
(C00), 78.9 (C00), 71.9 (C ), 67.2 (C00), 66.0 (C00), 62.4 (C ), 41.8 (C ),
40.1 (CH alkyl), 33.0 (2ꢄ CH2 alkyl), 26.2 (2ꢄ CH2 alkyl), 25.8 (CH2
alkyl). HRMS (ESI): m/z [M þ Na]þ calcd for C19H22N205Na:
381.1426, found: 381.1412.
References
[1] M. Ahmadian, D.E. Bergstrom, 5-Substituted nucleosides in Biochemistry and
Biotechnology, Modified Nucleosides in Biochemistry, Biotechnology and
Medicine, in: P. Herdewijn (Ed.), Wiley-VCH., Weinheim, 2008, pp. 249e276.
[2] E. De Clercq, Antiviral and antitumor activities of 5-substituted 20-deoxyur-
idines, Methods Find. Exp. Clin. Pharmacol. 2 (1980) 253e267 (review).
[3] S. Brakmann, S. Lobermann, High-density labelling of DNA: preparation and
characterization of the target material for single-molecule sequencing,
Angew. Chem., Int. Ed. 40 (2001) 1427e1429.
4.1.4.14. 5-[4-(Cyclopropyl)buta-1,3-diynyl]-20-deoxyuridine (31).
Yield 43%, 1H0NMR (400 MHz, MeOD)
d
8.41 (s, 1H, H6), 6.22 (t, 1H,
0
J ¼0 6.4 Hz, H1 ), 4.30 (dt, 1H, 6.1, 3.7 Hz, H3 ), 3.94 (q, J ¼ 3.2 Hz, 1H,
0
H4 ), 3.82 (dd, J ¼ 12.0 Hz, 3.0,1H, H5a ), 3.73 (dd, J ¼ 12.0 and 3.4 Hz,
[4] G. Giller, T. Tasara, B. Angerer, K. Mühlegger, M. Amacker, H. Winter, Incor-
poration of reporter molecule-labeled nucleotides by DNA polymerases. I.
0
0
1H, H5b ), 2.32 (ddd, J ¼ 13.6, 6.4, 3.8 Hz, 1H, H2a ), 2.27e2.18 (m, 1H,

228
O. Sari et al. / European Journal of Medicinal Chemistry 53 (2012) 220e228
Chemical synthesis of various reporter group-labeled 20-deoxyribonucleoside-
50-triphosphates, Nucleic Acids Res. 31 (2003) 2630e2635.
[5] I. Luyten, P. Herdewijn, Hybridization properties of base-modified oligonu-
cleotides within the double and triple helix motif, Eur. J. Med. Chem. 33 (1998)
515e576.
[6] T. Kottysch, C. Ahlborn, F. Brotzel, C. Richert, Stabilizing or destabilizing oli-
godeoxynucleotide duplexes containing single 20-deoxyuridine residues with
5-alkynyl substituents, Chem. Eur. J. 10 (2004) 4017e4028.
[15] Q. Xiao, R.T. Ranasinghe, AM.P. Tang, T. Brown, Naphthalenyl- and
anthracenyl-ethynyl dT analogues as base discriminating fluorescent nucle-
osides and intramolecular energy transfer donors in oligonucleotide probes,
Tetrahedron 63 (2007) 3483e3490.
[16] C. Glaser, Beiträge zur kenntniss des acetenylbenzols, Ber. Dtsch. Chem. Ges 2
(1869) 422e424.
[17] A.S. Hay, Oxidative coupling of acetylenes, J. Org. Chem. 27 (1962)
3320e3321.
[7] E. De Clercq, Discovery and development of BVDU (brivudin) as a therapeutic
for the treatment of herpes zoster, Biochem. Pharmacol. 68 (2004) 2301e2315
(review).
[18] G. Eglinton, A.R. Galbraith, Macrocyclic acetylenic compounds. Part I.
Cyclotetradeca-1: 3-diyne and related compounds, J. Chem. Soc. (1959)
889e896.
[8] C. McGuigan, A. Brancale, H. Barucki, S. Srinivasan, G. Jones, R. Pathirana,
A. Carangio, S. Blewett, G. Luoni, O. Bidet, A. Jukes, C. Jarvis, G. Andrei, R. Snoeck,
E. De Clercq, J. Balzarini, Furano pyrimidines as novel potent and selective anti-
VZV agents, Antivir. Chem. Chemother. 12 (2001) 77e89 (review).
[9] M. Kuwahara, Y. Takahata, A. Shoji, A.N. Ozaki, H. Ozaki, H. Sawai, Substrate
properties of C5-substituted pyrimidine 20-deoxynucleoside 50-triphosphates
for thermostable DNA polymerases during PCR, Bioorg. Med. Chem. Lett. 13
(2003) 3735e3738.
[10] D. Rai, M. Johar, T. Manning, B. Agrawal, D.Y. Kunimoto, R. Kumar, Design and
studies of novel 5-substituted alkynylpyrimidine nucleosides as potent
inhibitors of mycobacteria, J. Med. Chem. 48 (2005) 7012e7017.
[11] S. Meneni, I. Ott, C.D. Sergeant, A. Sniady, R. Gust, R. Dembinski, 5-Alkynyl-20-
deoxyuridines: chromatography-free synthesis and cytotoxicity evaluation
against human breast cancer cells, Bioorg. Med. Chem. 15 (2007) 3082e3088.
[12] I. Gillaizeau, Y. Saito, L.A. Agrofoglio, Palladium-assisted routes to nucleosides,
Chem. Rev. 103 (2003) 1875e1916 (review).
[19] E. Merkul, D. Urselmann, T.J.J. Müller, Consecutive one-pot Sonogashir-
aeGlaser coupling sequence e direct preparation of symmetrical diynes by
sequential Pd/Cu catalysis, Eur. J. Org. Chem. 2 (2011) 238e242.
[20] F. Yang, X. Cui, Y. Li, J. Zhang, G. Ren, Y. Wu, Cyclopalladated ferrocenylimines:
efficient catalysts for homocoupling and Sonogashira reaction of terminal
alkynes, Tetrahedron 63 (2007) 1963e1969.
[21] P. Siemsen, R.C. Livingston, F. Diederich, Acetylenic coupling: a powerful tool
in molecular construction, Angew. Chem., Int. Ed. 39 (2000) 2632 (review).
[22] P. Cadiot, W. Chodkiewicz, Coupling of Acetylenes, in: H.G. Viehe (Ed.),
Chemistry of Acetylenes, Marcel Dekker, New York, 1969, pp. 597e647.
[23] V. Escuret, V. Aucagne, N. Joubert, D. Durantel, K.L. Rapp, R.F. Schinazi,
F. Zoulim, L.A. Agrofoglio, Synthesis of 5-haloethynyl- and 5-(1,2-dihalo)
vinyluracil nucleosides: antiviral activity end cellular toxicity, Bioorg. Med.
Chem. 13 (2005) 6015e6024.
[24] T. Kurita, M. Abe, T. Maegawa, Y. Monguchi, H. Sajiki, Ligand- and base-free
synthesis of 1,3-diynes catalyzed by low loading of heterogeneous Pd/C and
CuI, Synlett 16 (2007) 2521e2524.
[25] W. Yin, C. He, M. Chen, H. Zhang, A. Lei, Nickel-catalyzed oxidative coupling
reactions of two different terminal alkynes using O2 as the oxidant at room
temperature: facile syntheses of unsymmetric 1,3-diynes, Org. Lett. 11 (2009)
709e712.
[13] A.L.K. Shi Shun, R.R. Tykwinski, Synthesis of naturally occurring polyynes,
Angew. Chem., Int. Ed. 45 (2006) 1034e1057.
[14] N. Minakawa, Y. Ono, A. Matsuda, A versatile modification of on-column oli-
godeoxynucleotides using a copper-catalyzed oxidative acetylenic coupling
reaction, J. Am. Chem. Soc. 125 (2003) 11545e11552.