Synthesis and anti-HCMV activity of 1-[ω-(phenoxy)alkyl]uracil derivatives and analogues thereof

Article abstract of DOI:10.1016/j.bmc.2013.05.009

HCMV infection represents a life-threatening condition for immunocompromised patients and newborn infants and novel anti-HCMV agents are clearly needed. In this regard, a series of 1-[ω-(phenoxy)alkyl]uracil derivatives were synthesized and examined for antiviral properties. Compounds 17, 20, 24 and 28 were found to exhibit highly specific and promising inhibitory activity against HCMV replication in HEL cell cultures with EC₅₀ values within 5.5-12 μM range. Further studies should be undertaken to elucidate the mechanism of action of these compounds and the structure-activity relationship for the linker region.

Full text of DOI:10.1016/j.bmc.2013.05.009

Accepted Manuscript
Synthesis and anti-HCMV activity of 1-[ω-(phenoxy)alkyl]uracil derivatives
and analogues thereof
Mikhail S. Novikov, Denis A. Babkov, Maria P. Paramonova, Anastasia L.
Khandazhinskaya, Alexander A. Ozerov, Alexander O. Chizhov, Graciela
Andrei, Robert Snoeck, Jan Balzarini, Katherine L. Seley-Radtke
PII:
S0968-0896(13)00434-3
DOI:
http://dx.doi.org/10.1016/j.bmc.2013.05.009
BMC 10835
Reference:
To appear in:
Bioorganic & Medicinal Chemistry
Received Date:
Revised Date:
Accepted Date:
8 April 2013
27 April 2013
7 May 2013
Please cite this article as: Novikov, M.S., Babkov, D.A., Paramonova, M.P., Khandazhinskaya, A.L., Ozerov, A.A.,
Chizhov, A.O., Andrei, G., Snoeck, R., Balzarini, J., Seley-Radtke, K.L., Synthesis and anti-HCMV activity of 1-
[ω-(phenoxy)alkyl]uracil derivatives and analogues thereof, Bioorganic & Medicinal Chemistry (2013), doi: http://
dx.doi.org/10.1016/j.bmc.2013.05.009
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Synthesis and Anti-HCMV Activity of 1-[ω-
(Phenoxy)alkyl]uracil Derivatives and
Analogues Thereof
Leave this area blank for abstract info.
Mikhail S. Novikova, Denis A. Babkova, Maria P. Paramonovaa, Anastasia L. Khandazhinskayab, Alexander A.
Ozerova, Alexander O. Chizhovc, Graciela Andreid, Robert Snoeckd, Jan Balzarinid, Katherine L. Seley-Radtkee*
aDepartment of Pharmaceutical & Toxicological Chemistry, Volgograd State Medical University, Pavshikh Bortsov Sq., 1,
Volgograd 400131, Russia;
bEngelhardt Institute of Molecular Biology, Russian Academy of Science, Vavilov Str., 32, Moscow 119991, Russia;
cZelinsky Institute of Organic Chemistry, Russian Academy of Science, Leninsky pr., 47, Moscow 119991, Russia;
dRega Institute for Medical Research, KU Leuven, Minderbroedersstraat 10, Leuven B-3000, Belgium;
eDepartment of Chemistry & Biochemistry, University of Maryland, Baltimore County, 1000 Hilltop Circle, Baltimore, MD
21250, USA.

Bioorganic & Medicinal Chemistry
journal homepage: www.elsevier.com
Synthesis and Anti-HCMV Activity of 1-[ω-(Phenoxy)alkyl]uracil Derivatives
and Analogues Thereof
Mikhail S. Novikova, Denis A. Babkova, Maria P. Paramonovaa, Anastasia L. Khandazhinskayab,
Alexander A. Ozerova, Alexander O. Chizhovc, Graciela Andreid, Robert Snoeckd, Jan Balzarinid, Katherine
L. Seley-Radtkee*
aDepartment of Pharmaceutical & Toxicological Chemistry, Volgograd State Medical University, Pavshikh Bortsov Sq., 1, Volgograd 400131, Russia;
bEngelhardt Institute of Molecular Biology, Russian Academy of Science, Vavilov Str., 32, Moscow 119991, Russia;
cZelinsky Institute of Organic Chemistry, Russian Academy of Science, Leninsky pr., 47, Moscow 119991, Russia;
dRega Institute for Medical Research, KU Leuven, Minderbroedersstraat 10, Leuven B-3000, Belgium;
eDepartment of Chemistry & Biochemistry, University of Maryland, Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250, USA.
ARTICLE INFO
ABSTRACT
Article history:
Received
Received in revised form
Accepted
HCMV infection represents a life-threatening condition for immunocompromised patients and
newborn infants and novel anti-HCMV agents are clearly needed. In this regard, the series of 1-
[ω-(phenoxy)alkyl]uracil derivatives were synthesized and examined for antiviral properties.
Compounds 17, 20, 24 and 28 were found to exhibit highly specific and promising inhibitory
activity against HCMV replication in HEL cell cultures with EC50 values within 5.5 - 12 μM
range. Further studies should be undertaken to elucidate the mechanism of action of these
compounds and the structure-activity relationship for the linker region.
Available online
Keywords:
Keyword_1
Keyword_2
Keyword_3
Keyword_4
Keyword_5
2013 Elsevier Ltd. All rights reserved.
however has been severely limited by their toxicity.18 In addition,
due to their inherent poor oral bioavailability, they must be
1. Introduction
Human cytomegalovirus (HCMV) belongs to the viral family
known as Herpesviridae, and is also known as human
herpesvirus-5 (HHV-5). Within Herpesviridae, HCMV belongs
to the Betaherpesvirinae subfamily.1 One characteristic feature of
the herpesviruses, including HCMV, is the ability to remain
latent within the body after infection.2 Latent HCMV is present in
approximately 90% of adults aged over 80 in the U.S.,3 and
typically goes unnoticed in healthy people, but can be life-
threatening for the immunocompromised person. In this regard,
HIV-infected persons,4,5 organ transplant recipients,6 or newborn
infants are all at high risk of infection. Transplacental HCMV
transmission can lead to congenital abnormalities and stillbirth,
thus represents one of the most common viral causes of birth
defects.7 HCMV infection may also cause mucoepidermoid
carcinoma and possibly other malignancies.8 Moreover, a number
of studies have revealed that HCMV is associated with
autoimmune diseases,9 atherosclerosis,10 coronary restenosis11,12
and increased risk of diabetes.13,14
administered intravenously to reach appropriate therapeutic
levels. The exception to this limitation is valganciclovir, the
orally administered prodrug for ganciclovir.19 Unfortunately, as is
typical for many nucleoside drugs, the development of drug-
resistant viral strains has emerged.20–22 Thus, there is an urgent
need for new and more effective anti-HCMV agents with
improved activity and pharmacokinetic profile.
Recently a new class of non-nucleoside HCMV polymerase
inhibitors has attracted interest.23 As a result of the BioChem
Pharm screening campaign, a series of 1,6-naphthiridine-based
HCMV inhibitors were identified. The highest activity was
exhibited by compound 1 (Figure 1) which possesses an ortho-
substituted benzyl moiety.24 Subsequent design modifications led
to the macrocyclic analogues represented by 2.25 These
compounds exhibited potent activity against HCMV and other
herpesviruses, however were accompanied by significant
cytotoxicity. Potent anti-HCMV and anti-VZV activity was also
noted for other heterocyclic-based inhibitors including
imidazo[1,2-a]pyridine derivatives 326–28 and benzothiadiazines
4.29,30 Other examples of broad-spectrum herpesvirus inhibitors
are represented by derivatives of fused heterocyclic systems
including the quinolones,31 thieno[2,3-b]-,32 thieno[3,2-b]-,33
furo[2,3-b]pyridines34 (5) and pyrrolo[3,2,1-ij]quinolones35 (6),
To date only three anti-HCMV agents, ganciclovir,15
cidofovir,16 and foscarnet,17 have been approved for clinical use.
They exhibit their effects by inhibiting the activity of the HCMV
polymerase, thus reducing viral replication in patients with
recognized HCMV infection symptoms. The use of these drugs

all developed by Pfizer (Figure 1). It should be noted that all of
these compounds possess a common structural feature: an N-
heterocycle linked to a benzene ring.
Scheme 2. Conditions: (a) aq. NaOH, EtOH, reflux, 8 hr.; (b)
SnCl2·H2O, EtOH, reflux, 3 hr.
2.2. Antiviral activity
The anti-HCMV properties of target compounds 9-30 were
evaluated against HCMV (AD-169 and Davis strains) in HEL
cell cultures and the results are shown in Table 1. Most of the
compounds proved to be inhibitory against HCMV. In examining
the structure activity relationship for the compounds, it appears
that the activity is strongly dependent upon the substituent at the
para-position of the benzene core. Polar groups proved
unfavorable and resulted in a loss of activity (compounds 18, 21,
29). Activity for alkyl substituted compounds increase with the
size of the substituent EC50 for H (9) ≈ Me (12) > i-Pr (14) > t-Bu
(15), however introduction of a bulky phenyl group led to poor
activity for 16, thus there appears to be a steric limit. The 4-
cyano- (17) and 4-bromo- (20) substituted compounds were
found to exhibit the highest activity, which was comparable to
ganciclovir. Interestingly, the 4-chloro-substituted compound 19
was essentially inactive. One possible rationale for these
observations could be complementary polar interactions between
the halogen- or cyano-substituents and an appropriate protein
functionality, e.g. a carbonyl group, since the oxygen-containing
compound 29 was inactive. Interestingly, this assumption is
partially supported by the fact that reduction of NO2-group in
inactive compound 21 led to 4-amino substituted compound 30
which exhibited slight anti-HCMV activity.
Figure 1. HCMV inhibitors.
Previously we have reported the synthesis and antiviral
properties for a number of different classes of nucleobase
derivatives. Specifically, the anti-HIV and anti-HCMV activity
for a series of 9-[2-(phenoxy)ethyl]- and 9-[2-(benzyloxy)ethyl]-
derivatives of adenine have been described.36 The 1-{[2-
(phenoxy)ethoxy]methyl}uracils were found to be moderately
active against HIV,37 while 1-[2-(2-benzoylphenoxy)ethyl]-
derivatives exhibited strong inhibitory properties.38 In addition,
several derivatives of 3-benzyl-1-(cinnamyl)uracil demonstrated
significant activity against HIV and HCMV replication in cell
culture.39 As a result, these observations provided strong impetus
to further explore the antiviral activity spectrum of 1-[ -
(phenoxy)alkyl]uracils, as well as to investigate the structural
requirements for the linker between the aromatic moieties, since
this scaffold is common to many known inhibitors of HCMV
polymerase.
Substitutions at position 5 of the uracil ring were also
examined. In that regard, while thymine analogue 24 was slightly
more active than uracil derivative 20, for halogen-substituted 22
and 23 activity was notably diminished. As a result, this
modification was not investigated further.
2. Results and discussion
2.1. Chemistry
A series of novel uracil derivatives was synthesized in a
similar manner as previously described by our group.38,40
Initially, 20 was selected to investigate optimal length of the
linker chain between the uracil and the benzene moieties.
Compounds 25, 26 and 27, containing three, four and six
methylenes respectively, were found to be inactive against
HCMV at subtoxic concentrations. In contrast, compound 28,
which has eight methylene units, demonstrated the same level of
activity as 20.
Condensation
of
equimolar
amounts
of
2,4-
bis(trimethylsilyl)pyrimidines 7 with bromides 8 was performed
at 160-170 C in the absence of solvent to afford target
compounds 9 – 28 in 73-88% yield (Scheme 1).
In addition, target compounds 9-30 were screened against a
large panel of other DNA and RNA viruses. No activity was
observed for HSV-1, HSV-2, VZV, Vaccinia virus, Vesicular
stomatitis virus, Coxsackie virus B4, Respiratory syncytial virus,
Influenza A virus H1N1 subtype, Influenza A virus H3N2
subtype, Influenza B virus, Para-influenza-3 virus, Reovirus-1,
Sindbis virus, Punta Toro virus, Feline Corona Virus, HIV-1 and
HIV-2. The only exception was noted for compound 13, which
Scheme 1. General approach for the synthesis of the target compounds.
Compound 29 was obtained in an 81% yield by alkaline
hydrolysis of 18 in refluxing water-ethanol for 8 hours (Scheme
2), while reduction of compound 21 was accomplished with
SnCl2 in mild acid-free conditions41,42 to produce the amino-
substituted compound 30 in 59% yield.
demonstrated
a
50%
reduction
of
HIV-1-induced
cytopathogenesis at 24 μM concentration with a 50% cytotoxic
concentration of 154 μM.

Table 1. Anti-HCMV activity of the synthesized compounds in HEL cell cultures.
Compd
R1
R2
n
mp
Yield
(%)
EC50 (µМ)a
Cytotoxicity (µМ)
(° C)
AD-169
Davis
Cell morphology
(MCC)b
>100
Cell growth
(CC50)c
>100
H
H
H
H
H
3
3
3
3
151-153
147-149
101-103
146-147
88
72
75
87
>100
>100
45.0
>20
>100
>100
55.0
>20
9
10
11
12
2-Me
3-Me
4-Me
>100
>100
>100
>100
100
>100
H
H
3,5-Me2
4-iPr
3
3
96-98
85
78
>100
25.0
>100
15.0
>100
100
>100
>100
13
14
139-140
H
H
H
H
H
H
H
Br
4-tBu
4-Ph
3
3
3
3
3
3
3
3
125-127
174-175
185-186
131-132
168-169
125-127
197-199
160-162
77
76
84
76
84
76
78
77
14.0
>20
8.9
12.0
54
100
100
40
15
16
17
18
19
20
21
22
ND
4-CN
5.5
100
100
n
4-CO2 Pr
45.0
>100
9.4
45.0
>100
12.0
>4
≥100
>100
100
>100
ND
4-Cl
4-Br
4-NO2
4-Br
>100
ND
>4
20
36.0
67.0
>100
>100
I
4-Br
4-Br
3
3
166-168
169-171
78
82
13.0
8.9
20.0
9.0
≥100
>100
>100
23
24
Me
100
H
H
4-Br
4-Br
1
2
179-181
143-144
79
80
>100
>20
>100
>20
>100
100
ND
ND
25
26
H
H
4-Br
4-Br
4
6
121-122
145-147
70
76
>4
>4
20
ND
27
28
12.0
8.9
≥100
>100
H
H
4-CO2H
4-NH2
3
3
225-227
190-192
81
59
>100
55.0
>100
55.0
>100
>100
ND
29
30
>100
Ganciclovir
Cidofovir
7.0
1.3
8.3
1.1
394
317
200
161
a Effective concentration required to reduce virus plaque formation by 50%. Virus input was 100 plaque forming units (PFU).
b Minimum cytotoxic concentration that causes a microscopically detectable alteration of cell morphology.
c Cytotoxic concentration required to reduce cell growth by 50%.
unless otherwise noted. Anhydrous DMF and isopropanol were
purchased from Sigma-Aldrich Co. Anhydrous acetone, 1,2-
dichloroethane, and ethyl acetate were obtained by distillation
over P2O5. NMR spectra were registered on a Bruker Avance 400
3. Conclusion
A series of novel 1-[ω-(phenoxy)alkyl]uracil derivatives was
synthesized and shown to have promising and highly specific
inhibitory properties against HCMV replication in cell culture.
Notably, the substitution pattern in the benzene core appears to
be of importance for activity. The EC50 values for the most active
compounds in the series (compounds 17, 20, 24 and 28) lie
within the range of 5.5 – 12.0 μM and are accompanied with low
cytotoxicity (CC50 100 µM). Additional efforts are currently
underway to elucidate the mechanism of action of these
compounds and further explore the structure-activity relationship
for the linker region.
1
spectrometer (400 MHz for H and 100 MHz for 13C) in DMSO-
d6 with tetramethylsilane as an internal standard. Data are
reported in the following order: multiplicity (br, broad; s, singlet;
d, doublet; dd, doublet of doublets; t, triplet; m, multiplet; q,
quartet; qu, quintet). TLC was performed on Merck TLC Silica
gel 60 F254 plates eluted with ethyl acetate or chloroform/MeOH
(10 : 1) and developed with UV-lamp VL-6.LC (France). Acros
Organics (Belgium) silica gel (Kieselgur 60-200 μm, 60A) was
used for column chromatography. Melting points were
determined in glass capillaries on a Mel-Temp 3.0 (Laboratory
Devices Inc., US). Yields refer to spectroscopically (1H and 13C
NMR) homogeneous materials. High resolution mass spectra
were measured on Bruker micrOTOF II instruments using
electrospray ionization (HRESIMS). The measurements were
4. Materials and methods
4.1. General
All reagents were obtained at highest grade available from
Sigma and Acros Organics and used without further purification

done in a positive ion mode (interface capillary voltage –4500 V)
in a mass range from m/z 50 to m/z 3000 Da; external or internal
calibration was done with ESI Tuning MixTM (Agilent
Technologies). A syringe injection was used for solutions in
acetonitrile (flow rate 3 µl/min). Nitrogen was applied as a dry
gas; interface temperature was set at 180 C. Bromides 8 were
synthesized as per known procedures.43,44
1.73 (2H, qu, J = 7.8 Hz, CH2), 2.21 (3H, s, CH3), 3.67 (2H, t, J =
7.3 Hz, NCH2), 3.88 (2H, t, J = 6.5 Hz, OCH2), 5.49 (1H, dd, J =
7.8 and 2 Hz, H-5), 6.73 (2H, d, J = 8.7 Hz, H-3’, H-5’), 7.01
(2H, d, J = 8.3 Hz, H-2’, H-6’), 7.47 (1H, d, J = 7.8 Hz, H-6),
11.07 (1H, s, NH). 13C NMR (DMSO-d6): 20.1, 22.5, 28.3, 28.4,
47.5, 67.1, 101.0, 114.1, 129.0, 129.6, 145.0, 150.9, 156.5, 163.8.
HRESIMS: found m/z 289.1542, calculated for C16H20N2O3
[M+H]+ 289.1547; m/z 311.1361, calcd for C16H20N2O3 [M+Na]+
311.1366.
4.2. Synthesis
4.2.1. General procedure for the synthesis of 1-[5-(phenoxy)-
pentyl]uracils (9 – 24).
4.2.1.5. 1-[5-(3,5-Dimethylphenoxy)pentyl]uracil (13). Yield
85%, mp 96-98 °C, Rf 0.58 (ethyl acetate); 1H NMR (DMSO-d6):
1.37 (2H, qu, J = 6.8 Hz, CH2), 1.62 (2H, qu, J = 7.2 Hz, CH2),
1.67 (2H, qu, J = 7.2 Hz, CH2), 2.21 (6H, s, CH3), 3.66 (2H, t, J =
7.1 Hz, NCH2), 3.88 (2H, t, J = 6.4 Hz, OCH2), 5.55 (1H, dd, J =
7.9 and 2.2 Hz, H-5), 6.51 (2H, s, H-2’, H-6’), 6.53 (1H, s, H-4’),
7.64 (1H, d, J = 7.8 Hz, H-6), 11.25 (1H, s, NH). 13C NMR
(DMSO-d6): 21.1, 22.5, 28.2, 28.4, 47.4, 66.9, 100.8, 112.1,
122.0, 138.6, 145.7, 151.0, 158.7, 163.8. HRESIMS: found m/z
303.1709, calculated for C17H22N2O3 [M+H]+ 303.1703; m/z
325.1526, calcd for C17H22N2O3 [M+Na]+ 325.1523.
A mixture of uracil, 5-bromouracil, 5-iodouracil or thymine
(5.60 mmol) and ammonium chloride (0.3 g, 5.60 mmol) in
HMDS (15 mL) was refluxed for 10 hr with exclusion of
moisture until a clear solution was obtained. Excess silylating
reagent was removed under vacuum. To the residual clear oil of
2,4-bis(trimethylsilyloxy)pyrimidine 7, an equimolar amount of
bromide 8 was added, the reaction mixture heated at 160-170 °C
for 1 hr, then the resulting amber oil was dissolved in 40 mL of
EtOAc, treated with 10 mL of i-PrOH and evaporated. The
residue was dissolved in 10 mL of CHCl3 and purified by column
chromatography, eluting with CHCl3/MeOH, 10:1. Subsequent
recrystallization from i-PrOH/DMF mixture provided the desired
products.
4.2.1.6. 1-[5-(4-i-Propylphenoxy)pentyl]uracil (14). Yield 78%,
1
mp 139-140 °C, Rf 0.50 (ethyl acetate); H NMR (DMSO-d6):
1.15 (6H, d, J = 7.0 Hz, CH3), 1.39 (2H, qu, J = 7.0 Hz, CH2),
1.63 (2H, qu, J = 7.6 Hz, CH2), 1.71 (2H, qu, J = 7.6 Hz, CH2),
2.80 (1H, m, J = 6.9 Hz, CH2), 3.67 (2H, t, J = 7.2 Hz, NCH2),
3.90 (2H, t, J = 6.4 Hz, OCH2), 5.55 (1H, dd, J = 7.8 and 2.2 Hz,
H = 5), 6.82 (1H, d, J = 8.7 Hz, H-3’, H-5’), 7.11 (2 H, d, J = 8.7
Hz, H-2’, H-6’), 7.65 (1H, d, J = 7.8 Hz, H-6), 11.24 (1H, d, J =
1.3 Hz, H-3). 13C NMR (DMSO-d6): 22.48, 24.13, 28.23, 28.37,
32.61, 39.11, 39.31, 39.52, 39.73, 39.94, 47.38, 67.10, 100.83,
114.20, 127.08, 140.24, 145.70, 150.97, 156.73, 163.79.
HRESIMS: found m/z 317.1863, calculated for C18H24N2O3
[M+H]+ 317.1860; m/z 339.1677, calcd for C18H24N2O3 [M+Na]+
339.1679.
4.2.1.1. 1-[5-(Phenoxy)pentyl]uracil (9). Yield 88%, mp 151-
153 °C, Rf 0.53 (ethyl acetate); 1H NMR (DMSO-d6): 1.39 (2H,
qu, J = 5.3 Hz, CH2), 1.63 (2H, qu, J = 7.2 Hz, CH2), 1.72 (2H,
qu, J = 7.2 Hz, CH2), 3.67 (2H, t, J = 7.2 Hz, NCH2), 3.93 (2H, t,
J = 6.5 Hz, OCH2), 5.55 (1H, dd, J = 7.7 and 2.1 Hz, H-5), 6.89-
6.92 (3H, m, aromatic H), 7.26 (2H, t, J = 8 Hz, H-3’, H-5’), 7.64
(1H, d, J = 7.8 Hz, H-6), 11.25 (1H, s, NH). 13C NMR (DMSO-
d6): 22.9, 28.6, 28.7, 47.8, 67.5, 101.2, 114.8, 120.8, 129.9,
146.2, 151.4, 159.3, 164.3. HRESIMS: found m/z 275.1394,
calculated for C15H18N2O3 [M+H]+ 275.1390; m/z 297.1214,
calcd for C15H18N2O3 [M+Na]+ 297.1210.
4.2.1.7. 1-[5-(4-tert-Butylphenoxy)pentyl]uracil (15). Yield
1
77%, mp 125-127 °C, Rf 0.50 (ethyl acetate); H NMR (DMSO-
4.2.1.2. 1-[5-(2-Methylphenoxy)pentyl]uracil (10). Yield 72%,
1
d6): 1.23 (9H, s, CH3), 1.38 (2H, qu, J = 7.2 Hz, CH2), 1.62 (2H,
qu, J = 7.2 Hz, CH2), 1.70 (2H, qu, J = 7.5 Hz, CH2), 3.67 (2H, t,
J = 7.1 Hz, NCH2), 3.90 (2H, t, J = 6.7 Hz, OCH2), 5.55 (1H, dd,
J = 7.8 and 2.2 Hz, H-5), 6.81 (2H, d, J = 8.9 Hz, H-3’, H-5’),
7.25 (2H, d, J = 8.9 Hz, H-2’, H-6’), 7.64 (1H, d, J = 7.8 Hz, H-
6), 11.25 (1H, s, NH). 13C NMR (DMSO-d6): 22.9, 28.7, 28.8,
31.8, 47.8, 67.5, 101.3, 114.3, 126.4, 142.9, 146.1, 151.4, 156.8,
164.3. HRESIMS: found m/z 331.2012, calculated for
C19H26N2O3 [M+H]+ 331.2016; m/z 353.1824, calcd for
C19H26N2O3 [M+Na]+ 353.1836.
mp 147-149 °C, Rf 0.51 (ethyl acetate); H NMR (DMSO-d6):
1.41 (2H, qu, J = 7 Hz, CH2), 1.64 (2H, qu, J = 7.3 Hz, CH2),
1.74 (2H, qu, J = 7.3 Hz, CH2), 2.12 (3H, s, CH3), 3.68 (2H, t, J =
7 Hz, NCH2), 3.92 (2H, t, J = 6.3 Hz, OCH2), 5.55 (1H, dd, J =
7.8 and 2 Hz, H-5), 6.80 (1H, t, J = 7.5 Hz, aromatic H), 6.87
(1H, d, J = 8 Hz, aromatic H), 7.09-7.13 (2H, m, aromatic H),
7.64 (1H, d, J = 7.9 Hz, H-6), 11.25 (1H, s, NH). 13C NMR
(DMSO-d6): 16.4, 22.9, 28.6, 28.8, 47.8, 67.5, 101.2, 111.6,
120.4, 126.9, 127.4, 130.8, 146.2, 151.4, 157.1, 164.3.
HRESIMS: found m/z 289.1541, calculated for C16H20N2O3
[M+H]+ 289.1547; m/z 311.1359, calcd for C16H20N2O3 [M+Na]+
311.1366.
4.2.1.8. 1-[5-(4-Phenylphenoxy)pentyl]uracil (16). Yield 76%,
1
mp 174-175 °C, Rf 0.54 (ethyl acetate); H NMR (DMSO-d6):
1.41 (2H, qu, J = 7.4 Hz, CH2), 1.64 (2H, qu, J = 7.4 Hz, CH2),
1.74 (2H, qu, J = 7.3 Hz, CH2), 3.68 (2H, t, J = 7.2 Hz, NCH2),
3.98 (2H, t, J = 6.4 Hz, OCH2), 5.56 (1H, dd, J = 7.8 and 2.2 Hz,
H-5), 7.00 (2H, d, J = 8.8 Hz, H-2’, H-6’), 7.30 (1H, t, J = 7.4
Hz, H-4’’), 7.42 (2H, t, J = 7.6 Hz, H-3’’, H-5’’), 7.54-7.63 (4H,
m, H-3’, H-5’, H-2’’, H-6’’), 7.66 (1H, d, J = 7.8 Hz, H-6), 11.26
(1H, s, H-3). 13C NMR (DMSO-d6): 22.44, 28.22, 28.30, 39.12,
39.33, 39.53, 39.74, 39.95, 47.38, 67.29, 100.84, 114.89, 126.16,
126.70, 127.75, 128.88, 132.44, 139.88, 145.73, 150.99, 158.30,
163.81. HRESIMS: found m/z 351.1708, calculated for
C21H22N2O3 [M+H]+ 351.1703; m/z 373.1526, calcd for
C21H22N2O3 [M+Na]+ 373.1523.
4.2.1.3. 1-[5-(3-Methylphenoxy)pentyl]uracil (11). Yield 75%,
mp 101-103 °C, Rf 0.49 (ethyl acetate); H NMR (DMSO-d6):
1
1.38 (2H, qu, J = 7 Hz, CH2), 1.64 (2H, qu, J = 7.3 Hz, CH2),
1.71 (2H, qu, J = 7.5 Hz, CH2), 2.25 (3H, s, CH3), 3.67 (2H, t, J =
7.2 Hz, NCH2), 3.90 (2H, t, J = 6.5 Hz, OCH2), 5.55 (1H, dd, J =
7.7 and 2 Hz, H-5), 6.68-6.72 (2H, m, aromatic H), 7.09-7.15
(2H, m, aromatic H), 7.63 (1H, d, J = 7.9 Hz, H-6), 11.23 (1H, s,
NH). 13C NMR (DMSO-d6): 21.1, 22.5, 28.2, 28.3, 47.4, 67.0,
100.8, 111.4, 115.1, 121.1, 129.2, 138.9, 145.7, 151.0, 158.7,
163.8. HRESIMS: found m/z 289.1552, calculated for
C16H20N2O3 [M+H]+ 289.1547; m/z 311.1372, calcd for
C16H20N2O3 [M+Na]+ 311.1366.
4.2.1.9. 1-[5-(4-Cyanophenoxy)pentyl]uracil (17). Yield 84%,
mp 185-186 °C, Rf 0.33 (chloroform/MeOH, 10 : 1); H NMR
4.2.1.4. 1-[5-(4-Methylphenoxy)pentyl]uracil (12). Yield 87%,
mp 146-147 °C, Rf 0.56 (ethyl acetate); H NMR (DMSO-d6):
1
1
(DMSO-d6): 1.40 (2H, qu, J = 7.9 Hz, CH2), 1.64 (2H, qu, J =
1.42 (2H, qu, J = 8 Hz, CH2), 1.66 (2H, qu, J = 7.1 Hz, CH2),

6.4 Hz, CH2), 1.75 (2H, qu, J = 7.4 Hz, CH2), 3.67 (2H, t, J = 7.2
Hz, NCH2), 4.05 (2H, t, J = 6.3 Hz, OCH2), 5.54 (1H, dd, J = 7.8
and 2.1 Hz, H-5), 7.07 (2H, d, J = 8.8 Hz, H-3’, H-5’), 7.62 (1H,
d, J = 7.9 Hz, H-6), 7.72 (2H, d, J = 9 Hz, H-2’, H-6’), 11.14
(1H, s, NH). 13C NMR (DMSO-d6): 22.3, 28.0, 28.1, 47.3, 67.9,
100.9, 102.8, 115.6, 119.1, 134.1, 145.6, 151.0, 162.2, 163.7.
HRESIMS: found m/z 300.1353, calculated for C16H17N3O3
[M+H]+ 300.1343; m/z 322.1163, calcd for C16H17N3O3 [M+Na]+
322.1162.
HRESIMS: found m/z 430.9603, calculated for C15H16Br2N2O3
[M+H]+ 430.9600; m/z 452.9426, calcd for C15H16Br2N2O3
[M+Na]+ 452.9420.
4.2.1.15. 1-[5-(4-Bromophenoxy)pentyl]-5-iodouracil (23).
1
Yield 78%, mp 166-168 °C, Rf 0.82 (ethyl acetate); H NMR
(DMSO-d6): 1.37 (2H, qu, J = 7.9 Hz, CH2), 1.62 (2H, qu, J =
7.4 Hz, CH2), 1.71 (2H, qu, J = 7.7 Hz, CH2), 3,67 (2H, t, J = 7.3,
NCH2), 3.93 (2H, t, J = 6.4 Hz, OCH2), 6.88 (2H, d, J = 9 Hz, H-
3’, H-5’), 7.40 (2H, d, J = 9 Hz, H-2’, H-6’), 8.20 (1H, s, H-6),
11.57 (1H, s, NH). 13C NMR (DMSO-d6): 25.7, 31.5, 31.6, 51.0,
70.9, 71.3, 115.1, 120.1, 135.4, 153.3, 154.0, 161.3, 164.4.
HRESIMS: found m/z 478.9445, calculated for C15H16BrIN2O3
[M+H]+ 478.9462; m/z 500.9274, calcd for C15H16BrIN2O3
[M+Na]+ 500.9281.
4.2.1.10.
1-[5-(4-n-Propoxycarbonylphenoxy)pentyl]uracil
(18). Yield 76%, mp 131-132 °C, Rf 0.35 (ethyl acetate); 1H
NMR (DMSO-d6): 0.94 (3H, t, J = 7.4 Hz, CH3) 1.39 (2H, qu, J
= 7.5 Hz, CH2) 1.57-1.81 (6H, m, CH2) 3.67 (2H, t, J = 7.1 Hz,
NCH2) 4.02 (2H, t, J = 6.4 Hz, OCH2) 4.17 (2H, t, J = 6.6 Hz,
OCH2) 5.55 (1H, dd, J = 7.8 and 2.1 Hz, H-5) 7.01 (2H, d, J =
8.8 Hz, H-2’, H-6’) 7.65 (1H, d, J = 7.8 Hz, H-6) 7.89 (2H, d, J =
8.7 Hz, H-3’, H-5’) 11.24 (1H, s, H-3). 13C NMR (CDCl3):
10.36, 21.69, 22.33, 28.11, 28.16, 39.04, 39.25, 39.46, 39.67,
39.88, 47.33, 65.72, 67.62, 100.82, 114.38, 121.96, 131.17,
145.72, 150.97, 162.50, 163.82, 165.48. HRESIMS: found m/z
361.1758, calculated for C19H24N2O5 [M+H]+ 361.1758; m/z
383.1568, calcd for C19H24N2O5 [M+Na]+ 383.1577.
4.2.1.16. 1-[5-(4-Bromophenoxy)pentyl]thymine (24). Yield
1
82%, mp 169-171 °C, Rf 0.60 (ethyl acetate); H NMR (DMSO-
d6): 1.38 (2H, qu, J = 8.1 Hz, CH2), 1.62 (2H, qu, J = 7.1 Hz,
CH2), 1.72 (2H, qu, J = 7.6 Hz, CH2), 1.75 (3Н, s, СН3), 3.63
(2H, t, J = 7.3 Hz, NCH2), 3.94 (2H, t, J = 6.4 Hz, OCH2), 6.89
(2H, d, J = 9 Hz, H-3’, H-5’), 7.42 (2H, d, J = 9.1 Hz, H-2’, H-
6’), 7.53 (1H, s, H-6), 11.21 (1H, s, NH). 13C NMR (DMSO-d6):
12.0, 22.4, 28.2, 47.0, 67.5, 108.4, 111.8, 116.7, 132.1, 141.5,
150.9, 157.9, 164.3. HRESIMS: found m/z 367.0638, calculated
for C16H19BrN2O3 [M+H]+ 367.0652; m/z 389.0460, calcd for
C16H19BrN2O3 [M+Na]+ 389.0471.
4.2.1.11. 1-[5-(4-Chlorophenoxy)pentyl]uracil (19). Yield 84%,
1
mp 168-169 °C, Rf 0.41 (ethyl acetate); H NMR (DMSO-d6):
1.38 (2H, qu, J = 6.1 Hz, CH2), 1.62 (2H, qu, J = 7.4 Hz, CH2),
1.71 (2H, qu, J = 7.6 Hz, CH2), 3.67 (2H, t, J = 7.3 Hz, NCH2),
3.92 (2H, t, J = 6.3 Hz, OCH2), 5.55 (1H, dd, J = 7.8 and 2.2 Hz,
H-5), 6.92 (2H, d, J = 8.7 Hz, H-3’, H-5’), 7.29 (2H, d, J = 8.7
Hz, H-2’, H-6’), 7.64 (1H, d, J = 7.9 Hz, H-6), 11.23 (1H, s,
NH). 13C NMR (DMSO-d6): 22.4, 28.2, 47.4, 67.6, 100.8, 116.1,
124.1, 129.2, 145.7, 151.0, 157.5, 163.8. HRESIMS: found m/z
309.1003, calculated for C15H17ClN2O3 [M+H]+ 309.1000; m/z
331.0816, calcd for C15H17ClN2O3 [M+Na]+ 331.0820.
4.2.1.17. 1-[3-(4-Bromophenoxy)propyl]uracil (25). Yield
1
79%, mp 179-181 °C, Rf 0.46 (ethyl acetate); H NMR (DMSO-
d6), 2.04 (2H, qu, J = 6.3 Hz, CH2), 3.83 (2H, t, J = 6.8 Hz,
NCH2), 3.99 (2H, t, J = 6.8 Hz, OCH2), 5.53 (4H, dd, J = 7.8 and
2.3 Hz, H-5), 6.87 (2H, d, J = 9.0 Hz, H-3’, H-5’), 7.43 (2H, d, J
= 8.9 Hz, H-2’, H-6’), 7.62 (1H, d, J = 7.8 Hz, H-6), 11.22 (1H,
s, H-3). 13C NMR (DMSO-d6): 27.86, 39.12, 39.33, 39.53,
39.74, 39.95, 45.35, 65.39, 100.85, 112.04, 116.74, 132.13,
145.85, 151.04, 157.68, 163.82. HRESIMS: found m/z 325.0193,
calculated for C13H13BrN2O3 [M+H]+ 325.0182; m/z 347.0007,
calcd for C13H13BrN2O3 [M+Na]+ 347.0002.
4.2.1.12. 1-[5-(4-Bromophenoxy)pentyl]uracil (20). Yield 76%,
1
mp 125-127 °C, Rf 0.50 (ethyl acetate); H NMR (DMSO-d6):
1.39 (2H, qu, J = 8.3 Hz, CH2), 1.62 (2H, qu, J = 7.2 Hz, CH2),
1.71 (2H, qu, J = 7.2 Hz, CH2), 3.66 (2H, t, J = 7.2 Hz, NCH2),
3.92 (2H, t, J = 6.3 Hz, OCH2), 5.55 (1H, dd, J = 7.8 and 2.2 Hz,
H-5), 6.87 (2H, d, J = 9 Hz, H-3’, H-5’), 7.41 (2H, d, J = 8.9 Hz,
H-2’, H-6’), 7.64 (1H, d, J = 7.8 Hz, H-6), 11.23 (1H, s, NH). 13C
NMR (DMSO-d6): 22.8, 28.6, 47.8, 68.0, 101.3, 112.2, 117.1,
132.5, 146.2, 151.4, 158.3, 164.3. HRESIMS: found m/z
375.0310, calculated for C15H17BrN2O3 [M+Na]+ m/z 375.0315.
4.2.1.18. 1-[4-(4-Bromophenoxy)butyl]uracil (26). Yield 81%,
1
mp 143-144 °C, Rf 0.40 (ethyl acetate); H NMR (DMSO-d6):
1.67-1.72 (4H, m, CH2), 3.72 (2H, t, J = 6.6 Hz, CH2), 3.97 (2H,
t, J = 5.8 Hz, CH2), 5.56 (1Н, dd, J = 7.8 and 2.2 Hz, Н-5), 6.90
(2H, d, J = 9.1 Hz, H-3’, H-5’), 7.43 (2H, d, J = 9 Hz, H-2’, H-
6’), 7.67 (1H, d, J = 7.9 Hz, H-6), 11.23 (1H, s, NH). 13C NMR
(DMSO-d6): 25.2, 25.6, 47.2, 67.4, 100.9, 111.9, 116.8, 132.1,
145.7, 151.0, 157.9, 163.8. HRESIMS: found m/z 361.0160,
calculated for C14H15BrN2O3 [M+Na]+ 361.0158.
4.2.1.13. 1-[5-(4-Nitrophenoxy)pentyl]uracil (21). Yield 78%,
1
mp 197-199 °C, Rf 0.43 (chloroform/MeOH, 10 : 1); H NMR
(DMSO-d6): 1.39 (2H, qu, J = 7.6 Hz, CH2) 1.64 (2H, qu, J =
7.4 Hz, CH2) 1.76 (2H, qu, J = 7.2 Hz, CH2) 3.67 (2H, t, J = 7.2
Hz, NCH2) 4.10 (2H, t, J = 6.4 Hz, OCH2) 5.54 (1H, dd, J = 7.8
and 2.2 Hz, H-5) 7.11 (2H, d, J = 9.2 Hz, H-2’, H-6’) 7.65 (1H,
d, J = 7.8 Hz, H-6) 8.17 (2H, d, J = 9.2 Hz, H-3’, H-5’) 11.21
(1H, s, H-3). 13C NMR (DMSO-d6): 22.25, 27.98, 28.11, 39.12,
39.33, 39.53, 39.74, 39.95, 47.30, 68.42, 100.83, 114.97, 125.89,
145.71, 150.98, 163.78, 164.00. HRESIMS: found m/z 320.1234,
calculated for C15H17N3O5 [M+H]+ 320.1241; m/z 342.1048,
calcd for C15H17N3O5 [M+Na]+ 342.1060.
4.2.1.19. 1-[6-(4-Bromophenoxy)hexyl]uracil (27). Yield 74%,
mp 121-122 °C, Rf 0.51 (ethyl acetate); H NMR (DMSO-d6):
1
1.30 (2H, qu, J = 6.9 Hz, CH2), 1.42 (2Н, qu, J = 6.8 Hz, СН2),
1.59 (2Н, qu, J = 7.1 Hz, СН2), 1.70 (2Н, qu, J = 6.8 Hz, СН2),
3.65 (2H, t, J = 7.2 Hz, CH2), 3.93 (2H, t, J = 6.5 Hz, CH2), 5.53
(1Н, dd, J = 7.8 and 2.2 Hz, Н-5), 6.89 (2H, d, J = 9 Hz, H-3’, H-
5’), 7.42 (2H, d, J = 8.9 Hz, H-2’, H-6’), 7.64 (1H, d, J = 7.8 Hz,
H-6), 11.19 (1H, s, NH). 13C NMR (DMSO-d6): 25.1, 25.5,
28.4, 47.467.7, 100.8, 111.8, 116.8, 132.1, 145.7, 151.0, 158.0,
163.8. HRESIMS: found m/z 389.0465, calculated for
C16H19BrN2O3 [M+Na]+ 389.0471.
4.2.1.14. 1-[5-(4-Bromophenoxy)pentyl]-5-bromouracil (22).
1
Yield 77%, mp 160-162 °C, Rf 0.50 (ethyl acetate); H NMR
4.2.1.20. 1-[8-(4-Bromophenoxy)octyl]uracil (28). Yield 79%,
(DMSO-d6): 1.38 (2H, qu, J = 8.1 Hz, CH2), 1.64 (2H, qu, J =
7.2 Hz, CH2), 1.72 (2H, qu, J = 7.4 Hz, CH2), 3.69 (2H, t, J = 7.4
Hz, NCH2), 3.94 (2H, t, J = 6.5 Hz, OCH2), 6.89 (2H, d, J = 8.9
Hz, H-3’, H-5’), 7.42 (2H, d, J = 9 Hz, H-2’, H-6’), 8.24 (1H, s,
H-6), 11.74 (1H, s, NH). 13C NMR (DMSO-d6): 22.3, 28.1,
47.8, 67.5, 94.6, 111.8, 116.7, 132.1, 145.4, 150.4, 157.9, 159.7.
1
mp 145-147 °C, Rf 0.54 (ethyl acetate); H NMR (DMSO-d6):
1.16-1.43 (8H, m, CH2), 1.56 (2H, qu, J = 7.1 Hz, CH2), 1.67
(2H, qu, J = 7.0 Hz, CH2), 3.63 (2H, t, J = 7.2 Hz, NCH2), 3.92
(2H, t, J = 6.5 Hz, OCH2), 5.54 (1H, dd, J = 7.8 and 2.2 Hz, H-5),
6.88 (2H, d, J = 8.9 Hz, H-3’, H-5’), 7.41 (2H, d, J = 8.9 Hz, H-
2’, H-6’), 7.64 (1H, d, J = 7.8 Hz, H-6), 11.22 (1 H, s, H-3). 13C

NMR (DMSO-d6): 25.40, 25.76, 28.43, 28.54, 28.59, 28.64,
39.12, 39.33, 39.53, 39.74, 39.95, 47.44, 67.73, 100.78, 111.72,
116.71, 132.10, 145.71, 150.95, 157.98, 163.78. HRESIMS:
found m/z 395.0964, calculated for C18H23BrN2O3 [M+H]+
395.0965; m/z 417.0781, calcd for C18H23BrN2O3 [M+Na]+
417.0784.
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 formation by 50%.
4.3.2. Anti-HIV Activity Assays. Inhibition of HIV-1(IIIB)- and
HIV-2(ROD)-induced cytopathicity in CEM cell cultures was
measured in microtiter 96-well plates containing 3 × 105 CEM
cells/mL infected with 100 CCID50 of HIV per milliliter and
containing appropriate dilutions of the test compounds. After 4−5
days of incubation at 37 °C in a CO2-controlled humidified
atmosphere, CEM giant (syncytium) cell formation was
examined microscopically. The EC50 (50% effective
concentration) was defined as the compound concentration
required to inhibit HIV-induced giant cell formation by 50%.
4.2.2. 4-[5-(2,4-Dioxo-3,4-dihydropyrimidin-1(2H)-yl)pentyl-
oxy]benzoic acid (29).
A solution of 1-[5-(4-n-propoxy-
carbonylphenoxy)pentyl]uracil 18 (1.60 g, 4.44 mmol) in 40 mL
of EtOH and 20 mL of 2.5% aqeous NaOH was refluxed for 8 hr
and evaporated. The residue was cooled to rt, treated dropwise
with 2% HCl and stored in a refrigerator overnight. The resulting
precipitate was filtered and recrystallized from aqeous EtOH to
yield 1.15 g of 25 as white crystals. Yield 81%, mp 225-227 °C,
1
Rf 0.69 (chloroform/MeOH, 10:1); H NMR (DMSO-d6): 1.38
(2H, qu, J = 7.3 Hz, CH2), 1.63 (2H, qu, J = 7.3 Hz, CH2), 1.74
(2H, qu, J = 7.3 Hz, CH2), 3.66 (2H, t, J = 7.2 Hz, NCH2), 4.01
(2H, t, J = 6.3 Hz, OCH2), 5.54 (1H, d, J = 7.8 Hz, H-5), 6.98
(2H, d, J = 8.8 Hz, H-2’, H-6’), 7.64 (1H, d, J = 7.8 Hz, H-6),
7.87 (2H, d, J = 8.8 Hz, H-3’, H-5’), 11.22 (1H, br. s., H-3). 13C
NMR (DMSO-d6): 22.35, 28.13, 28.16, 39.07, 39.28, 39.49,
39.69, 39.90, 47.34, 67.57, 100.82, 114.21, 122.88, 131.37,
145.71, 150.97, 162.25, 163.80, 167.08. HRESIMS: found m/z
341.1108, calculated for C16H18N2O5 [M+Na]+ 341.1108; m/z
363.0928, calculated for C16H17NaN2O5 [M+Na]+ 363.0927.
4.3.3. Cytostatic Activity Assays. All assays were performed in
96-well microtiter plates. To each well were added (5−7.5) × 104
tumor cells and a given amount of the test compound. The cells
were allowed to proliferate for 48 h (murine leukemia L1210
cells) or 72 h (human lymphocytic CEM and human cervix
carcinoma HeLa cells) at 37 °C in a humidified CO2-controlled
atmosphere. At the end of the incubation period, the cells were
counted in
a Coulter counter. The IC50 (50% inhibitory
concentration) was defined as the concentration of the compound
that inhibited cell proliferation by 50%.
4.2.3. 1-[5-(4-Aminophenoxy)pentyl]uracil (30). To the
solution of 1-[5-(4-nitrophenoxy)pentyl]uracil 21 (1.17 g, 3.36
mmol) in 20 ml of abs. EtOH SnCl2·2H2O (4.2 g, 18.61 mmol)
was added. Resulting mixture was refluxed for 3 hr and poured
into 200 ml of cold distilled water. Resulting precipitate was
filtered off, air-dried and recrystallized from i-PrOH/DMF 2:1
mixture to yield 0.63 g of 26 as small yellow crystals. Yield 59%,
Acknowledgments
This work was supported by grant of Russian Foundation For
Basic Research (13-04-01391). The antiviral work was supported
by KU Leuven (GOA 10/14). The authors thank Lies Van den
Heurck, Anita Camps, Steven Carmans, Leentje Persoons, Frieda
De Meyer, Leen Ingels and Lizette van Berckelaer for their help
in performing the antiviral and cytostatic assays.
1
mp 190-192 °C, Rf 0.55 (chloroform/MeOH, 10 : 1); H NMR
(DMSO-d6): 1.36 (2H, qu, J = 7.4 Hz, CH2), 1.51-1.74 (4H, m,
CH2), 3.66 (2H, t, J = 7.1 Hz, NCH2), 3.80 (2H, t, J = 6.4 Hz,
OCH2), 4.53 (2H, br. s., NH2), 5.53 (1H, d, J = 7.8 Hz, H-5), 6.50
(2H, d, J = 8.7 Hz, H-2’, H-6’), 6.62 (2H, d, J = 8.8 Hz, H-3’, H-
5’), 7.63 (1H, d, J = 7.8 Hz, H-6), 11.17 (1H, br. s., H-3). 13C
NMR (DMSO-d6): 22.50, 28.23, 28.52, 39.15, 39.37, 39.58,
39.78, 40.00, 47.41, 67.77, 100.80, 115.01, 115.42, 142.32,
145.71, 150.97, 163.77. HRESIMS: found m/z 290.1507,
calculated for C15H19N3O3 [M+H]+ 290.1499; m/z 312.1325,
calcd for C15H19N3O3 [M+Na]+ 312.1319.
Supplementary data
Supplementary data (NMR spectra for all of the synthesized
compounds) associated with this article can be found online.
References and notes
1. Sherris Medical Microbiology; Ryan, K. J.; Ray, C. G., Eds.;
4th ed.; McGraw Hill., 2004; p. 556.
2. Reeves, M.; Sinclair, J. In Human Cytomegalovirus; Springer
Berlin Heidelberg, 2008; pp. 297–313.
3. Staras, S. A. S.; Dollard, S. C.; Radford, K. W.; Flanders, W.
D.; Pass, R. F.; Cannon, M. J. Clin. Infect. Dis. 2006, 43,
1143–51.
4. Baroco, A. L.; Oldfield, E. C. Curr. Gastroent. Rep. 2008,
10, 409–416.
5. Jackson, S. E.; Mason, G. M.; Wills, M. R. Virus Res. 2011,
157, 151–60.
6. Nashan, B.; Gaston, R.; Emery, V.; Säemann, M. D.; Mueller,
N. J.; Couzi, L.; Dantal, J.; Shihab, F.; Mulgaonkar, S.; Seun
Kim, Y.; Brennan, D. C. Transplantation 2012, 93, 1075–85.
7. Pereira, L. J. Infect. Dis. 2011, 203, 1510–2.
8. Melnick, M.; Sedghizadeh, P. P.; Allen, C. M.; Jaskoll, T.
Exp. Mol. Pathol. 2012, 92, 118–25.
9. Toyoda-Akui, M.; Yokomori, H.; Kaneko, F.; Shimizu, Y.;
Takeuchi, H.; Tahara, K.; Yoshida, H.; Kondo, H.; Motoori,
T.; Ohbu, M.; Oda, M.; Hibi, T. Intern. J. Gen. Med. 2011, 4,
397–402.
4.3. Biological assays
4.3.1. Antiviral Activity Assays other than HIV. The
compounds were evaluated against the following viruses: herpes
simplex virus type 1 (HSV-1) strain KOS, thymidine kinase-
deficient (TK−) HSV-1 KOS strain resistant to ACV (ACVr),
herpes simplex virus type 2 (HSV-2) strains Lyons and G, human
cytomegalovirus (HCMV) (strains AD-169 and Davis), varicella-
zoster virus (strains OKA and YS), vaccinia virus Lederle strain,
respiratory syncytial virus (RSV) strain Long, vesicular
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virus A (subtypes H1N1, H3N2), influenza virus B, Reovirus-1,
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