Hamites R.A., Schuster A.M., Loginova S.ya., Borisevich S.V. Of faldina V.N., Koval'chuk A.V.

Experimental data on the effectiveness Of arbidola with respect to the agent of atypical pneumonia (SARS) in vivo and in vitro.

(virusological center OF NII - SCIENTIFIC RESEARCH INSTITUTE microbiology MO RF)

The search for the effective medical means of preventive maintenance and treatment of "atypical pneumonia" (SARS) is vital problem. It is at present known that the inhibiting action on the culture of the cells of virus SARS render interferons (a, b), and also ribavirin in the high concentrations (500-5000 g/ml).

Purpose of a study. To study in vitro and in vivo the effectiveness of aribidola (PRIVATELY HELD COMPANY Of "masterlek") and its derived - mezilata arbidola ((NPF "SINBIAS FARMA", g.Donetsk, the Ukraine, the supplier PRIVATELY HELD COMPANY Of "masterlek") with respect to agent SARS.

Material and the methods of study.

During the first stage was investigated the antiviral activity of preparations in vitro to the virus of atypical pneumonia, the strain of sodas (statement about the delivery of patent RF from 17.12.2003 g. VKh в„– 2003136244 for the strain of the sodas of the virus of the heavy sharp respiratory syndrome of kind Coronavi- Russian, intended for developing of means and methods of biological protection). Were used the interwoven cultures of the cells of the kidneys of the green marmosets Gmk-ah-1(d) and Vero Ye'; SPEV (culture of the cells of the kidneys of the embryo of pig), semisynthetic medium (P) in the solution Of khenksa, which contains 7,5% and 2 % the sera of large livestock as the growth medium and the medium of maintenance. In preliminary studies according to the estimation of the reproduction of virus SARS (strain of sodas) in the interwoven cultures of cells and possibility to form negative colonies, under the solid agar coating, established that the virus is multiplied well in Gmk- AN -1(.d), Vero E6 and SPEV forms negative colonies under the agar only in the culture of cells Vero E6.

The antiviral action of preparations was studied in vivo (second stage). With planning of a study the doses of preparations for the oral and parenteral introduction and the diagram of their application are substantiated. Taking into account that during the infection of different it is specific animals (white mice, guinea pigs, rabbits, white rats, hens, pigs, monkeys) disease in the majority of the cases flows inapparantno and/or without their loss, for the estimation of the effectiveness of preparations were used the Syrian hamsters, orally infected with virus SARS.

The study of the flow of infection in Syrian hamsters showed that the virus produces statistically reliable (P<0,05) morphological changes in the lungs of the infected animals in comparison with the intact. Observation of the infected animals was conducted during 21 days, in this case the level of the accumulation of agent in the lungs was checked, were evaluated laboratory indices at the initial period of infection (2 days), on the peak of infection (4 days) and to 6 and 10 days.

Reduction in the caption of virus in the lungs to 1,5 lg and more, the degree of the manifestation of the defeat of lungs in comparison with the control group of animals, the positive dynamics of laboratory indices were criteria the effectivenesses of the action of preparations. The toxicity of preparations was evaluated on the uninfected Syrian hamsters.

Results of a study.

First stage.

Effectiveness of preparations to the infection of the cultures of the cells:

Arbidol and mezilat of arbidola proved to be effective with their introduction into the culture of cells Vero Ye' within the different periods to the infection with the initially established dose of virus. more effective proved to be mezilat of arbidola, in the concentration five times of less than arbidol.

Results proved to be positive and with the use of the low infecting doses (on the average of 0,0003 BOE/kl). Effect from mezilata of arbidola was achieved in the dose more than 3 times of less than arbidola. The maximum effectiveness of mezilata of arbidola on the suppression of the reproduction of virus composed 90% with its introduction into nutrient medium 2 h prior to infection.

Effectiveness of preparations at the moment and after the infection of the cultures of the cells:

In the culture of cells Gmk- AN-1(.D) at the moment and after their infection (infecting dose of virus - on the average of 0,0003 BOE/kl, period of incubation - 48 h) the preparations suppressed the reproduction of virus SARS (arbidol into the concentration 50 g/ml - to 2,3 lg, mezilat of arbidola in the concentration 25 g/ml - to 2,4 lg). The coefficient of inhibition for arbidola at the moment of infection composed 98,8%, after infection - 99,9 %, for mezilata of arbidola after infection - 99,6 %.

In the culture of cells SPEV (infecting dose of virus - on the average of 0,003 BOE/kl, the period of incubation - 72 h.) the antiviral effectiveness of arbidola composed 99,7%, mezilata of arbidola - 99,2%.

Both preparations proved to be most effective with their introduction into nutrient medium of the culture of the cells 2 hours after their infection.

With the combination arbidola or mezilata of arbidola ribavirinom was observed neither additive nor sinergidnogo effect.

Second stage.

Toxic effect in Syrian hamsters with the oral the introduction of arbidola at the dose from 60 to 200 mg/kg of mass and in the parenteral introduction at the dose of 'mg/kg was not observed.

With the experimental form of SARS the oral introduction of arbidola according to therapeutic and prophylactic diagram at the dose 60 mg/kg of mass suppressed the reproduction of virus in the lungs to 1,4-4,3 lg. In this case morphological changes considerably decreased, were reduced gravity of the flow of process, manifestation of changes in the laboratory indices.

During the application of arbidola at the dose of 60 mg/kg by 2 and 4 days after infection the level of the accumulation of virus was reduced to 93,8%. To 10 days the reproduction of virus in practice completely was suppressed both during the oral and parenteral application of preparation.

Therapeutic effectiveness on the suppression of the reproduction of virus in the lungs on the peak of infection was less expressed than therapeutic and prophylactic.

Mezilat of arbidola in the therapeutic and prophylactic diagram of application suppressed the multiplication of virus in the lungs to 2,5-3,1 lg, in this case reliably (P<0,05) was reduced the gravity of process.

Conclusion.

The represented data about the effectiveness of arbidola and mezilata of arbidola with the experimental form of SARS testify about the expediency of further actions for the substantiation of their application with "atypical pneumonia".