Studies » Antiviral Effect of EnergoIod Preparation
Antiviral Effect of EnergoIod Preparation
The range of major antiviral preparations used in clinical practice for threatment of viral diseases is indicative of insufficiency of chemotherapeutic preparations selectively suppressing virus-specific processes in cells. Antiviral preparations are still less effective in comparison with broad spectrum antibiotics. Moreover most of them affect relatively small number of viruses. Therapeutic effect of antiviral preparations is inferior to their preventive effect and use of most of them is restricted due to their toxicity. In the light of these facts study of antiviral activity of new preparation EnergoIod - water solution of complex iodine compounds with low-molecular and high-molecular synthetic and natural compounds seemed to be reasonable.
In the course of studies antiviral activity of this preparation against influenza virus (IV) of type A, strain А/А ichi 2/68 (H3N2) - a representative of orthomyxoviruses, virus of human herpetic fever of the second serotype (family of herpes viruses) and virus of encephalomyocarditis - VEM (family of picornaviruses) was studied.
Materials and methods:
Preparations. Original studied preparation was used in the form of water solutions with dilution rate from 1/5 to 1/80 prepared ex tempore. Toxicity of the preparation was studied in cell culture, mice and chicken embryos (CE).
Official antiviral preparations - remantadin and acyclovir were used as positive controls in experiments with mice and CE.
Viruses. Experiments were conducted with influenza virus (IV) of type A, virus of herpetic fever of the second serotype (VHF) and virus of mouse encephalomyocarditis - (VEM).
Cell cultures. For intensification and accumulation of viruses and study of antiviral activity continuous line of human laryngeal adenocarcinoma Hep-2 and primary fibroblastic culture were used.
CE-FCE. Cells were cultivated in Eagle’s medium with glutamine and bovine serum (Hep-2) and in 199 medium with serum (FCE).
Animals. 160 outbred white mice with weight of 12-14 g. were used. Infection was administered intranasally and intraperitoneally. For accumulation of VHF and VEM - infection was administered intracerebrally.
CE. 140 CE aged 10-12 days were used. Viruses and the preparation were administered to allantoic cavity and chorioallantoic membrane.
Methods of study:
Titration of viruses in cell cultures by cytopathic effect. Titre determination by Kerber’s method.
Titration of viruses in mice using Reed-Muench method.
Titration of viruses in chicken embryos (CE).
Hemagglutination reaction (HAR).
Antiviral effect of EnergoIod preparation with various doses of viruses was determined by the rate of reproduction of viruses in cell culture and CE, by effectiveness of protection of mice against experimental viral infections (criteria - lethality, reproduction of viruses in lungs, duration of incubation period and average length of life).
This preparation was used in various experimental conditions: administration before infection, after infection and simultaneously with infection.
Obtained results were processed with application of statistical methods.
Analysis of obtained data confirms antiviral activity of this preparation against viruses of various families of “vira” kingdom - viruses of influenza A/A or 2/68 H3N2 (orthomyxoviruses), virus of herpetic fever of the second serotype (P-herpes viruses) and virus of encephalomyocarditis (VEM).
Studies were conducted in 3 phases: in vitro, in developing CE and in vivo - in mice.
The preparation was nontoxic for cell culture in concentrations of 1/20, 1/40, 1/80, for CE (1/10, 1/20, 1/40, 1/80) and mice - 1/5, 1/10, 1/20, 1/40. Antiviral effect of the preparation was shown in all phases with simultaneous, prior and following intraperitoneal administration, however effect was dose dependant and was optimal with prior administration.
Antiviral effect of the preparation was expressed by positive inhibition of reproduction of viruses in cell cultures and CE, protection against development of experimental influenza infection and experimental encephalomyocarditis in mice, positive reduction of animals’ mortality, prevention of reproduction of viruses in lungs, prolongation of incubation period and average length of life of mice.
Our studies were not aimed at examination of the mechanisms of antiviral action of the preparation using methods of molecular biology, however a range of facts established in studies of the preparation’s interaction with viruses at the cell level confirms the preparation’s virucidal effect and ability to disturb early stages of viral reproduction - adsorption, penetration into cells and probably deproteinization - undressing of viruses.
The fact that experiments with mixture of VEM (dose 2.5x105 TCD 50) with the preparation (dilution rate 1/20) matured at room temperature did not cause cytopathic effect of viruses in cells is indicative the preparation’s ability to neutralize viruses in the specified dose, meanwhile control VEM in the same dose caused total degeneration of layer in 24 hours. This data are indicative of the preparation’s viricidal action, which does not exclude its ability to affect cells sensitive to viruses, prevent adsorption and penetration of viruses into cells. This is confirmed by the previously described experiments where prior administration of the preparation followed by its thorough pumping out and washing of Hep-2 cells inhibited replication of virus for 2-3 log in comparison with control. Irrespective of withdrawal of the preparation it could cause changes in physicochemical properties of cells membranes during adsorption, in particular their charge, which in its turn could result in disturbance of virus’s contact with specific cell receptors. This is the mechanism of action of many surface-active preparations, in particular ethonium. Remantadin accumulated in cells membranes changes their charge and pH and prevents undressing of viruses - one of early stages of viral infection.
So there are good grounds for believing that the studied preparation disturbs early stages of infectious cycle and prevents expression of viral genome and further stages of reproduction of viruses - transcription, translation, replication of viral RNA and DNA, assembly of virions and their coming out of cells.
The main active component of the studied preparation is iodine and in this connection it would be good to mention other antiviral preparations containing iodine such as ethane, iodine antipyrilamide, 5-iodine-2- deoxyuridine and others.
Data obtained in in vitro and in vivo experiments with regard to antiviral effect of EnergoIod and its nontoxicity, as well as good solubility in water, ability to neutralize viruses outside cells and disturb early stages of viral reproduction, presence of both therapeutic and preventive action and rather broad-spectrum activity allow to regard it as an effective preparation for chemotherapy of viral infections.