New composition of tungsten has a broad range of antiviral activity

Abstract

The water-based combination of two inorganic chemical compounds such as sodium tungstate dihydrate-Na₂WO₄ × 2H₂O and Aluminum sulfate octadecahydrate-Al₂ (SO₄) 3 × 18H₂O that we have conditionally named 'Vomifal' has a broad antiviral activity in various DNA and RNA viruses, including Human Herpes Virus (HHV), African Swine Fever Virus (ASFV), Vaccinia Virus (VV), Hepatitis C Virus (HCV), Foot and Mouth Disease Virus (FMDV), Influenza A virus (A/Aichi/2/68 (H3N2)). In vitro and In vivo assays in several tissue cultures as well as in laboratory animals, conformed 'Vomifal' has a very low toxicity and the antiviral properties partially are due to its ability to induce gamma-IFN. Based on the results obtained, we can assume the presence of at least two mechanisms of the antiviral action of the studied drug. First or early stage - an unknown mechanism, possibly related to the effect on cellular receptors. Second or late stage - main antiviral properties probably associated with an interferonogenic effect.

Keywords: Hepatitis C virus; influenza; interferon; picornaviridae; poxviridae.

Figure 1.

Cytotoxicity of different concentrations of “Vomifal” in cultural medium. Values represent mean from three independent experiments. Data have been analyzed by Student's t-test. Significant differences compared to control are denoted by * p < 0.05–0.01.

Figure 2.

Antiviral action of “Vomifal” in vitro. (a) ‘Vomifal’ reduced HCV viral RNA level in Huh7-Lunet cells. Cells were pretreated at control or indicated the concentration of “Vomifal” subsequently infected with 4 × 10³-HCV particles. At 5 dpi viral RNA level was measured by rtPCR. IFN was used as inhibitory control. (b) Effect of ‘Vomifal’ on FMDV-A. BHK-21 cells were infected with FMDV-A virus in the absence or presence of “Vomifal” at 0.2% of stack solution in culture medium, at the indicated time post infection cell lysate was harvested and virus titer was determined in BHK-21 cells. (c) ASFV viral titer in primary porcine alveolar macrophages. Control or ‘Vomifal’ pretreated AMs were infected with ASFV at conc. of 10⁴ HAD₅₀/mL at indicated time post infection cell lysates were harvested and titer was determined in primary porcine AM. (d) Vaccinia virus plaque formation was reduced with ‘Vomifal’. BSC40 cells were pretreated with medium alone (a, b) or with medium containing 1:20 dilution of ‘Vomifal’ (c, d) for 1hr at 37 °C cells were then infected with wild type WR-VV of 100 PFU (a, c) or 50 PFU (b, d) cultured in medium containing ‘Vomifal’ fixed and stained with crystal violet at 2 day pi. * Data were analyzed by Student's t-test Significant comp’red to control (p < 0.05–p < 0.01); ** tendency (p < 0.1).

Figure 3.

Antiviral effect of “Vomifal” in vivo. (a) HHV1 infection of mice. 8 mice/group were fed by water or the indicated concentration of “Vomifal” followed by intra-brain virus inoculation Average life expectancy white bars and the percentage of survivals black line is shown. Data were analyzed by Student's t-test. (b) HHV1 viral t'ter in brain tissue. On the 1^st, 3^rd, 5^th and 7^th day po^st in^fecte^d animal^s with or without ‘Vomifal’ were sacrificed and virus titer in the brain was determined. Data were analyzed by Student's t-test. (c), (d) General’zation of FMDV-A virus on Guinea pigs, with/without ‘Vomifal’. Control (black bar), ‘Vomifal’ (white bar), 1 h prior to subcutaneous inoculation animals were fed (8 animals per group) by 0.2 mL/g (c) or 1 mL/g (d) of ‘Vomifal’. Data were analyzed by Mann-Whitney U-test. (e) Influenza A A/Aichi/2/68 (H3N2)) virus infection of mice. 20 mice/group were fed by water orally the indicated concentration of “Vomifal” followed by injections virus. Average life expectancy dark gray and percentage of lethality (light gray) is shown. Data were analyzed by Mann-Whitney U-test. * Significance (p < 0.05–p < 0.01); ** tendency (p < 0.1).

Figure 4.

Gamma-IFN induction by “Vomifal”. (a) Gamma-IFN induction by “Vomifal” at in vitro conditions. Primary human mononuclear cells were control treated (black) or treated with PHA (white), (0.2% solution in cell medium) ‘Vomifal (gray) or both (light gray). At the indicated time of post treatment, gamma-IFN induction of supernatant was measured using classical ELISA assay. (b) Gamma-IFN induction by “Vomifal” in in vivo conditions on murine model. 1st group - single per os administration of “Vomifal” in dose 430 µg/kg. 2nd group - single per os administration of “Vomifal” in dose 250 µg/kg. 3rd group - every day per os administration of “Vomifal” in single dose 150 µg/kg (total in five days 750 µg/kg). Data were analyzed by Student's t-test. * Significance (p < 0.05–p < 0.01).