1. DEVELOPMENT OF EFFECTIVE ANTIVIRAL AGENTS OF A NEW TYPE
Professor
Oleg Shadyro
Belarusian State University
Department of Chemistry, Minsk, Belarus

2. The main goal of the study was the development of antivital agents based on the substances capable of regulating various types of free radical reactions.

3. Advantages of the product / technology
Some time ago, we have developed an antiviral product Butaminophen that has proven to be effective against herpetic injuries of various types.
Advantages of the product / technology
The Product
is an effective anti-herpetic agent,
particularly against strains resistant to acyclovir,
it possesses also wound-healing, anti-inflammatory and antipyrotic action.
The Technology
is simple and easy to put into practice,
is a low-cost manufacturing process,
the starting raw material is readily available.

4. A general scheme depicting synthetic pathways to obtain some sterically hindered aminophenol derivatives

5. Membrane structure
Phospholipids O P X C

6. Lipid peroxidation process

7. Free-radical fragmentation of cardiolipin
Shadyro O.I., Yurkova I.L., Kisel M.A., Brede O., Arnhold J. Radiation-induced fragmentation of cardiolipin in a model membrane. International Journal of Radiation Biology, 2004, 80,
Shadyro O.I., Yurkova I.L., Kisel M.A., Brede O., Arnhold J. Radiation-induced free-radical transformations of Phospholipids: MALDI-TOF MS study. Chemistry and Physics of Lipids, 2004, 132,

8. Free-radical fragmentation of cerebrosides
Shadyro O.I., Yurkova I.L., Kisel M.A., Brede O., Arnhold J. Formation of phosphatidic acid, ceramide and diglyceride on radiolysis of lipids: identification by MALDI-TOF mass spectrometry. Free Radical Biology & Medicine, 2004, 36,
Shadyro O.I., Yurkova I.L., Kisel M.A., Arnhold J. Free-radical fragmentation of galactocerebrosides: a MALDI-TOF mass spectrometry study. Chemistry and Physics of Lipids, 2005, 134,
Shadyro O.I., Yurkova I.L., Kisel M.A., Arnhold J. Iron-mediated free-radical formation of signaling lipids in a model system. Chemistry and Physics of Lipids, 2005, 137,

9. A new approach to the regulation of free-radical processes in biosystems has been proposed
Diphenol and aminophenol derivatives were found to be capable of regulating free-radical transformations occurring in bioorganic compounds with participation of both oxygen-centered (oxidation) and carbon-centered (fragmentation) radicals.
Shadyro O.I. et al. Quinones as free-radical fragmentation inhibitors in biologically important molecules. Free Rad. Res., 2002, 36,
Shadyro O.I., Murase H., Kagiya T. et al. Effects of phenolic compounds on reactions involving various organic radicals. Free Rad. Res., 2003, 37,
Shadyro O.I. et al. Reactions of arylamine and aminophenol derivatives, and riboflavin with organic radicals. Free Rad. Res., 2004, 38,

10. Percent inhibition produced by aminophenols in reactions involving various radicals
Test compounds
Structure
>CHOO
>CH
>CHOH
N-1 28 81 92
N-2 48 44 45
N-3 34
N-4 49 41
N-5 33 77 78
N-6 17 76
N-7 58 83 80
N-8
1,2
4,6
0,8
N-9
9,1
8,3
2,3

11. Concentration range, M
Effective concentrations of aminophenols inhibiting the zymosan-stimulated production of ROS by macrophages
Test
compounds
Structure
Concentration range, M
EC50, M
EC90, M
N-1
0.06
0.65
N-2
No inhibition
N-3
N-4
N-5
9.8
> 10
N-6
N-7
N-9

12. Antiviral properties of the test compounds in a cell culture infected with HSV
Structure
MNTC, M
EC50 (I95*), M
EC90 (I95*), M
N-1
113.2
87.3
(214.935.3)
288.2
(709.9117.2)
N-2
379.7
8.5
(10.56.9)
14.8
(18.212.1)
N-3
720.9
38.2
(41.335.3)
64.5
(69.659.5)
N-4
686.2
8.6
(10.37.2)
14.1
(17.211.7)
N-5
336.7
23.0
(56.49.4)
169.4
(316.290.9)
N-6
643.1
30.9
(37.025.7)
83.0
(99.469.1)
N-7
611.6
18.0
(22.414.5)
41.9
(52.333.6)
N-8
1444.0
798.0
(1053.8604.3)
1960.5
(2588.81484.8)
N-9
722.0
255.2
(569.2114.4)
623.8
(1373.9283.3)
*I95 — is confidence interval at 95 % probability.

13. Wounds in places of vesicle formation
Antiviral properties of the test compounds in mice infected with skin herpes
Normal ear
Erythema
Erythema and vesicles
Wounds in places of vesicle formation

14. Higher antiviral activity against herpes viruses
Chemico-pharmacological advantages of compound N-2 as compared to Butaminophen®
Lower toxicity
Higher antiviral activity against herpes viruses
Higher chemical stability

15. Antiviral activity of compound N-2 against influenza A/FPV/Rostok (H7N1) virus in chicken embryo cell culture
Compound
code
Concentration, M
Titer of virus,
lg PFU/ml
EC50, M
MNTC/ EC50
N-2
759
380
190 48 24 12
4.2
4.5
4.6
4.7
5.4
5.5
43.6
17.4

16. Antiviral activity of compound N-12 against HIV-1 in a cell culture
Compound code
Concentration, M
Percent of infected cells
EC50, M
MNTC/ EC50
N-12
876
221 55 14 12 27 32 41 96
7.9
111

17. Conclusions:
The obtained data indicate that sterically hindered aminophenol derivatives possess antiviral properties and hence may be regarded as a novel class of antiviral agents.
Among the compounds tested, the most pronounced antiviral properties were found for N-acyl and N-aryl dertivatives of sterically hindered o-aminophenol which were able to interact with various organic radicals while displaying low reactivity towards reactive oxygen species.