SYNTHESIS AND ANTIVIRAL ACTIVITY
561
O-Benzyloxime of 5-formyl-3',5'-di-O-acetyl-2'-
O-Allyloxime of 5-formyl-2'-deoxyuridine (Xd);
yield: 80%; UV (H2O): λmax 293.5 nm; 1H NMR (D2O):
deoxyuridine (IXc); yield 92%; UV (H2O): λmax
1
293.9 nm; H NMR (CDCl3): 9.85 (1 H, br. s, 3-NH), 9.03 (0.26 H, s, 5-CH= syn), 8.11 (0.74 H, s, H6 anti),
7.89 (0.74 H, s, 5-CH= anti), 7.28 (0.26 H, s, H6 syn),
6.13 (0.26 H, t, J1'2' 7.1, H1' syn), 6.09 (0.74 H, t, J1'2' 7.0,
H1' anti), 5.93–5.84 (2 H, m, H2'), 5.23 (0.74 H, d,
2J2'',3''a 1.6, H3''a anti), 5.19 (0.26 H, d, 2J2'',3''a 1.6, H3''a
8.99 (0.35 H, s, 5-CH=, syn), 8.15 (0.65 H, s, H6, anti),
8.07 [0.65 H, s, 5-CH=, anti), 7.50 (0.35 H, s, H6, syn),
3
7.42–7.25 (5 H, m, C6H5), 6.30 (0.65 H, dd, J1'2' 8.4,
3J1'2' 6.2, H1' anti), 6.19 (0.35 H, dd, 3J1'2' 7.8, 3J1'2' 5.9,
H1' syn), 5.24 (0.35 H, m, H3' syn), 5.18 (0.65 H, m,
H3' anti), 5.10 (2 × 0.65 H, s, CH2C6H5 anti), 4.95 (2 ×
0.35 H, s, CH2C6H5 syn), 4.39–4.28 (3 × 0.65 H, m, H5',
H4' anti), 4.13 (0.35 H, m, H4' syn), 3.88 (0.35 H, dd,
syn), 5.16 (0.74 H, d, 2J2'',3''a 8.6, H3''b anti), 5.14 (0.26
H, d, 2J2'',3''a 8.5, H3''b syn), 4.54 (0.26 H, d, 3J1'',2'' 5.7,
3
H1'' syn), 4.47 (0.74 H, d, J1'',2'' 5.9, H1'' anti), 4.31
[0.74 H, m, H3' (anti), 4.27 [0.26 H, m, H3' syn), 3.89
(1 H, m, H4'), 3.70 [0.74 H, dd, 2J5'a,5'b 11.5, 3J5'a,4' 3.1,
H5'a anti), 3.65 [0.74 H, dd, 3J5'a,4' 4.7, H5'b anti), 3.63
3
2J5'a,5'b 12.1, J5'a,4' 4.7, H5'a syn), 3.73 (0.35 H, dd,
3
2J5'a,5'b 12.1, J5'a,4' 4.1, H5'b, syn), 2.58–2.45 (1 H, m,
2
H2'b), 2.25–2.18 (1 H, m, H2'a), 2.10 (3H, s, CH3), and
1.98 (3H, s, CH3).
[0.26 H, m, H5'a syn), 3.54 (0.26 H, dd, J5'a,5'b 11.2,
3J5'a,4' 4.2, H5'b syn), and 2.38–2.13 (2 H, m, H2').
O-Allyloxime of 5-formyl-3',5'-di-O-acetyl-2'-
deoxyuridine (IXd); yield 88%; UV (H2): λmax 293.7
Experiments on Cell Cultures
1
nm; H NMR (CDCl3): 9.69 (1 H, br. s, 3-NH), 9.03
A reference strain HSV-1/L2 was obtained from the
State Collection of Viruses of the Ivanovsky Institute of
Virology, Russian Academy of Medical Sciences, Rus-
sia. The strains were passaged in 1 : 1 mixture of the
Eagle and 199 media with the addition of 2% fetal calf
serum. The Vero cell culture (kidney cells of African
green marmosets) clone E6 was used in the work. The
cells were cultured in the Eagle’s MEM medium (the
Institute of Poliomyelitis and Viral Encephalitides,
Russian Academy of Medical Sciences, Moscow) con-
taining 7% fetal calf serum (PanEco, Moscow).
The antiherpetic activity was determined according
to the ability of compounds tested to inhibit the growth
of the virus-induced cytopathogenic effect by 50%
using the procedure described in [7]. Acyclovir [9-(2-
hydroxyethoxymethyl)guanine, Glaxo Wellcome, UK]
was used as a reference compound.
Cytotoxicity of the compounds was determined on
uninfected cultures by the procedure based on the abil-
ity of dead cells to be stained with Trypan Blue dye [7].
CD50 was taken as a concentration of compound, at
which no more than 50% cells died after 72-h incuba-
tion with the compound tested [11].
The antipox effect of the compounds synthesized
was studied with five viral strains: human smallpox,
monkeypox, cowpox, mousepox, and vaccinia viruses.
They were obtained from the State Collection of
Viruses of the Vector State Center of Virology and Bio-
technology. Vero and MK2 cell cultures were used for
the activity and toxicity studies. The antiviral effect was
evaluated as the viability of the virus-infected cells
incubated with the compounds studied.
(0.38 H, s, 5-CH=, syn), 8.10 (0.62 H, s, H6 anti), 8.07
[0.62 H, s, 5-CH=, anti), 7.54 (0.38 H, s, H6, syn), 6.32
(0.62 H, dd, J1'2'a 8.4, J1'2'b 5.6, H1' anti), 6.24 (0.38 H,
dd, J1'2'a 7.2, J1'2'b 6.2, H1' syn), 6.02–5.88 (1 H, m,
H2''], 5.38–5.10 (3 H, m, H3', H3''), 4.65 (0.38 H, d,
3
3J1'',2'' 5.6, H1'' syn), 4.56 (0.62 H, d, J1'',2'' 5.6, H1''
anti), 4.33 (2 × 0.62 H, m, H5' anti), 4.29 (2 × 0.38 H,
m, H5' syn, 0.62 I, m, H4' anti), 4.24 (0.38 H, m, H4'
syn), 2.58–2.45 (1 H, m, H2'a), 2.25–2.18 (1 H, m,
H2'b), 2.12 (3 H, s, CH3), and 1.97 (3 H, s, CH3).
Oxime of 5-formyl-2'-deoxyuridine (Xa); yield
83%; UV (H2O): λmax 289.5 nm; 1H NMR (D2O): 9.12
(0.37 H, s, 5-CH=, syn), 8.07 [0.63 H, s, 5-CH=, anti),
7.86 (0.63 H, s, H6, anti), 7.31 (0.37 H, s, H6, syn), 6.16
(0.37 H, t, J1'2' 6.5, H1' syn), 6.11 (0.63 H, t, J1'2' 6.2, H1'
anti), 4.33–4.22 (1 H, m, H3'), 3.94–3.85 (1 H, m, H4'),
3.73–3.55 (2 H, m, H5'), and 2.35–2.15 (2 H, m, H2').
O-Methyloxime of 5-formyl-2'-deoxyuridine
(Xb); yield 86%; UV (H2O): λmax 293.1 nm; 1H NMR
(D2O): 9.08 (0.35 H, s, 5-CH= syn), 8.24 (0.65 H, s, H6
anti), 8.00 (0.65 H, s, CH= anti), 7.42 (0.35 H, s, H6
syn), 6.28–6.20 (1 H, s, H1'), 4.47–4.41 (1 H, m, H3'),
4.06–4.01 (1 H, m, H4'), 3.82 (3 H, m, CH3), 3.77–3.73
(2 H, m, H5'), and 2.46–2.33 (2 H, m, H2').
O-Benzyloxime of 5-formyl-2'-deoxyuridine
(Xc); yield 78%; UV (H2O): λmax 293.6 nm; 1H NMR
(D2O): 9.17 (0.33 H, s, 5-CH= syn), 8.44 (0.67 H, s, H6,
anti), 8.01 (0.67 H, s, 5-CH= anti), 7.45–7.32 (0.35 H
+ 5 H, m, H6 syn and C6H5), 6.30–6.22 (1 H, m, H1'),
5.23 (2 × 0.33 H, s, CH2C6H5 syn), 5.13 (2 × 0.67 H, s,
CH2C6H5 anti), 4.40 (0.67 H, m, H3' anti), 4.19 (0.33
H, m, H3' syn), 3.95 (1 H, m, H4' anti), 3.88 (0.33 H, m,
H4' syn), 3.80 (0.67 H, dd, 2J5'a,5'b 11.8, 3J5'a,4' 3.4, H5'a
ACKNOWLEDGMENTS
2
3
anti), 3.74 (0.67 H, dd, J5'a,5'b 11.8, J5'a,4' 3.8, H5'b
We are grateful to V.L. Andronova (Ivanovskii Insti-
anti), 3.46 (2 × 0.33 H, m, H5'a syn), 2.38–2.25 (2 × tute of Virology, Russian Academy of Medical Sci-
0.67 H + 0.33 H, m, H2' anti, 0.33 H, m, H2'a syn), ences) for the experiments on the antiherpetic activity
2.18–2.02 (0.33 H, m, H2'a syn).
of the compounds synthesized.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 31 No. 6 2005