RU2446802C2 - Tabletted antiviral agent and method for preparing it - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to pharmaceutics, specifically to a tabletted antiviral agent. The agent contains, wt %: as an active substance - 2-methyl-6-nitro-1,2,4-triazolo[5,1-c]-1,2,4-triazin-7-one sodium salt, dehydrate (60.0-90.0), as an excipient - potato starch and/or lactose (1.0-20.0), as an aerating agent - low-substituted hydroxypropyl cellulose and/or microcrystalline cellulose (0.5-20.0), as antifriction substances - talc (0.2-2.5) and aerosil (0.2-3.0), as a greasing ingredient - magnesium stearate (0.2-3.0), as an agent forming a protective coating - hypromellose (1.0-10.0). The declared agent exhibits virus-inhibiting activity with respect to avian influenza virus, Rift Valley fever viruses, parainfluenza virus, Aujeszky's disease virus, western equine sleeping sickness virus, Venezuelan sleeping sickness virus, respiratory syncytial virus. What is also presented is a method for preparing said tabletted agent.

EFFECT: group of inventions provides the extended range of antiviral preparations due to preparing the tabletted agent having manifested virus-inhibiting action and improved organoleptic properties.

10 cl, 2 dwg, 6 tbl, 4 ex

Description

The claimed invention relates to the chemical and pharmaceutical industry, in particular to the compositions of drugs with antiviral effects, mainly produced in tablets, as well as to methods for their preparation.

A significant number of antiviral drugs are known from the prior art, such as arbidol, amixin (tilorone), ribavirin, acyclovir and others, most of which are available in solid dosage forms, namely tablets and capsules (see, for example, Mashkovsky M.D. Medicines. - 15th ed. M.: New Wave LLC. - 2006).

From literature data it is known that gelatin capsules have a number of disadvantages, for example, high sensitivity to moisture, therefore certain conditions must be met to preserve the drug. In addition, gelatin capsules are a favorable medium for the propagation of microorganisms, to prevent exposure to which preservatives, such as nipagin, sorbic acid and others, must be added to the composition of the drug (see, for example, Chueshov V.I., Chernov M.Yu. Industrial technology of medicines. - X .: MTK-Kniga, 2002. - v.2).

The tablet dosage form has a number of technological and pharmacological advantages over other forms [see, eg, Ershov F.I. Chizhov N.P. Tazulakhova E.B. Antiviral agents. St. Petersburg, 1993; Chizhov N.P. Anikin V.B. Romantsov M.G.// "Doctor". - 1993. - N3]:

- full mechanization of the manufacturing process, providing high performance, cleanliness and hygiene of the tablets;

- accuracy of dosage of drugs;

- long-term preservation of medicinal substances in a compressed state;

- masking unpleasant organoleptic properties (taste, smell). This is achieved by applying sugar shells; cocoa; Chocolate

- the possibility of a combination of medicinal substances incompatible in their physicochemical properties in other forms;

- localization of the action of the drug. This property is achieved by applying shells of a special composition, soluble in an acidic (stomach) or alkaline (intestine) environment;

- prolonged action of drugs;

- regulation of the sequential absorption of several drugs from the tablet at certain intervals - the creation of multilayer tablets.

The closest in technical essence the solution to the claimed composition is an antiviral agent containing amixin as an active substance and excipients selected as a prototype (see RF patent for the invention No. 2141314 "Antiviral drug" Amiksin "and method for its preparation", filing date 05/12/1998, published on 11/20/1999). As a filler, basic magnesium carbonate or magnesium oxide is used. As a humectant, polyvinylpyrrolidone or polyvinyl alcohol. For dusting, calcium stearate or aluminum stearate is used. The shell contains aerosil, wax, polyvinylpyrrolidone, sucrose or lactose, liquid paraffin or castor oil, tartrazine, basic magnesium carbonate or magnesium oxide, titanium dioxide. Sucrose and lactose give the drug a pleasant taste. Glossy mixture gives the drug a pleasant appearance. The ratio of all of the above components, wt.%:

Amixin - 30.5-33.6

Basic magnesium carbonate or magnesium oxide - 17.8-18.2

Polyvinylpyrrolidone (PVP) or polyvinyl alcohol (PVA) - 0.48-0.52

Magnesium or Calcium Stearate - 0.48-0.52

The shell is, on average, 51.3% of the total weight of the tablet and consists of the following components, wt.%:

Aerosil - 0.62-0.67

Wax - 0.04-0.05

Basic magnesium carbonate or magnesium oxide or calcium carbonate - 16.9-17.5

Vaseline or castor oil - 0.04-0.05

Polyvinylpyrrolidone - 0.78-0.89

Sucrose or lactose - 27.5-28.8

Talc - 0.12-0.13

Tartrazine - 0.03-0.04

Titanium Dioxide - 1.25-1.36.

The disadvantages of the known composition are the use of polyvinylpyrrolidone as the filler, the main magnesium carbonate and humectant, as well as the application of a multicomponent coating complicates the composition of the tablet, which significantly lengthens the process and makes the entire production process more expensive.

The technical result, to which the claimed invention is directed, is to obtain a tabletted drug with improved organoleptic properties, pharmacological safety, provided that it fully complies with the requirements of the Pharmacopoeia, as well as optimizing the method of producing the drug and expanding the range of drugs with antiviral effects.

The claimed technical result is achieved by the fact that in an antiviral agent in tablets containing an active substance, physiologically acceptable auxiliary additives, namely a filler, a disintegrant, an anti-friction substance, a lubricant and a protective coating, according to the invention, 2-methyl- sodium salt is used as an active substance 6-nitro-1,2,4-triazolo [5,1-s] -1,2,4-triazin-7-one, dihydrate, as filler - potato starch and / or lactose, as a disintegrant - hydroxypropyl cellulose with low grade enyu substitution and / or microcrystalline cellulose; as antifriction substances - talc and aerosil; magnesium stearate as a lubricating component; as a substance for the formation of a protective shell - hypromellose in the following ratio of components, wt.%:

Sodium salt of 2-methyl-6-nitro-1,2,4-triazolo [5.1-s] -1,2,4-triazin-7-one dihydrate 60.0-90.0 Potato starch and / or lactose 1,0-20,0 Low Substitution Hydroxypropyl Cellulose and / or microcrystalline cellulose 0.5-20.0 Aerosil 0.2-3.0 Magnesium stearate 0.2-3.0 Talc 0.2-2.5 Hypromellose 1.0-10.0

Tablets of an antiviral agent are made weighing 0.300-0.330 g, coated with a protective coating.

The inventive antiviral agent has a wide spectrum of virus-inhibiting activity, including in relation to bird flu virus, Rift virus, parainfluenza virus, Aujeszky's disease virus, Western horse encephalomyelitis, Venezuelan horse encephalomyelitis, respiratory syncytial virus.

In the proposed composition, the ratio of the active component and the target additives, obtained experimentally, is optimal and provides high-quality tablets. The proposed composition of the antiviral agent provides a good therapeutic effect. Starch and / or lactose affect the rate of dissolution, while an increase in starch and / or lactose (more than 20% wt.%) Leads to a decrease in tablet strength and an increase in tablet abrasion. In this case, the use of starch and / or lactose improves the dissolution process of the tablet, thereby easily releasing the active component, which ensures a high absorption rate of the active component in the gastrointestinal tract. The introduction of microcrystalline cellulose and / or hydroxypropyl cellulose with a low degree of substitution in the selected amount provides good hardness of the tablets and reduces their fragility. In addition to the above properties of microcrystalline cellulose and / or hydroxypropyl cellulose with a low degree of substitution, they prevent the action of moisture, which may be in other auxiliary substances, on the active component. Amixin tablets contain basic magnesium carbonate as a filler, which exhibits the maximum cementation effect in tablets. In contrast to the proposed composition of the tablets, the above effect is practically absent. The introduction of hydroxypropyl cellulose with a low degree of substitution and / or microcrystalline cellulose in the selected amount provides excellent tablet strength, due to the fibrous structure, the tablet is able to resist damage in all directions. Therefore, only less than 1% of all tablets have a “capping” effect (peeling of the top of the tablet). This, in turn, reduces the economic losses of production due to a decrease in the number of substandard tablets. Magnesium stearate was chosen as a lubricating component, since its lubricity is highest (unlike calcium sterate in amixin tablets). A decrease in the amount of dusting substances — talc, aerosil and magnesium sterate — (less than 0.2 wt.%) Worsens the sliding properties of the granulate during the tabletting process, which leads to the low-tech tableting process. The claimed tool does not have the ability to direct pressing and therefore, to obtain a solid dosage form of this active substance, a careful selection of excipients is necessary.

Amixin tablets are coated with a multicomponent sugar coating, which complicates the entire process. Coating the tablets of the proposed composition with a film coating leads to a more rapid release of the active component from the tablet and, accordingly, to a more rapid onset of action of the drug; in addition, fewer excipients that enter the sugar shell will enter the human body.

As can be seen from the foregoing, only the proposed ranges of ratios of the ingredients obtained experimentally are optimal and allow you to get a high-quality antiviral agent in the form of tablets that meets the requirements for drugs in all respects: in appearance, disintegration, dissolution, uniformity of dosage and other indicators. This dosage form is stable during storage and has a shelf life of more than 3 years.

The claimed ratio of known components is optimal, as it allows you to get a higher result compared with known drugs, in particular, expands the arsenal of drugs with a wide spectrum of action.

Technical solutions that coincide with the totality of the essential features of the claimed invention have not been identified, which allows us to conclude that the claimed invention meets such a patentability condition as “novelty”.

The claimed essential features that predetermine the receipt of the specified technical result, do not explicitly follow from the prior art, which allows us to conclude that the claimed invention meets such a patentability condition as "inventive step".

The patentability condition “industrial applicability” is confirmed by the example of a specific application.

Declares a method of obtaining an antiviral agent in tablets.

The prior art method for producing tablets of an antiviral agent, comprising wetting a mixture of amixin with an excipient, followed by wet granulation, drying, dry granulation, dusting, tabletting, coating by suspension pelleting. Amixin substance is mixed with a filler - basic magnesium carbonate. Then add a humectant in the form of an alcoholic solution of polyvinylpyrrolidone prepared previously. The prepared mixture is passed through a granulator with a hole diameter of 2 mm. The obtained granulate is dried and fed to dry granulation with a hole diameter of 1.5 mm Dusting is carried out with calcium stearate. Powdered granules are fed to a tablet press. The resulting tablets are fed into a pelletizing boiler for sequential application of sugar syrup, suspension and colored syrup, respectively. The final step is to apply a glossy mixture to give shine to the tablets (see RF patent for the invention No. 2141314 "Amiksin antiviral drug and method for its preparation", filing date 05/12/1998, published 11/20/1999).

The technical result to which the claimed invention is directed is to optimize the technology for producing a broad-action antiviral agent with good organoleptic properties and pharmacological safety, corresponding to the indicators of the pharmacopeia article of the enterprise.

This result is achieved by the fact that in the method of producing an antiviral preparation in tablets, carried out using the active substance, physiologically acceptable auxiliary additives: filler, disintegrant, antifriction substance, lubricant and protective coating and including moisturizing, subsequent wet granulation, drying, dry granulation, dusting , tableting and film coating, in which, when mixing the active substance, filler, baking powder, granulating the resulting mass, after which the moistened mass is first wiped through a perforated device, dried, and drying is carried out to a residual moisture content of not more than 0.5%; then secondary rubbing is carried out through a perforated device, after which the granulate obtained is dusted with a mixture of talc, aerosil and magnesium stearate, then tableted and coated with a protective coating, while the sodium salt of 2-methyl-6-nitro-1,2,4 is used as the active substance triazolo [5,1-c] -1,2,4-triazin-7-one, dihydrate, as filler, potato starch and / or lactose, as a disintegrant, low substitution hydroxypropyl cellulose and / or microcrystalline cellulose; as antifriction substances - talc and aerosil; as a lubricating component - magnesium stearate; as a substance for the formation of a protective shell - hypromellose, in the following ratio of components, wt.%:

Sodium salt of 2-methyl-6-nitro-1,2,4-triazolo [5.1-s] -1,2,4-triazin-7-one dihydrate 60.0-90.0 Potato starch and / or lactose 1,0-20,0 Low Substitution Hydroxypropyl Cellulose and / or microcrystalline cellulose 0.5-20.0 Aerosil 0.2-3.0 Magnesium stearate 0.2-3.0 Talc 0.2-2.5 Hypromellose 1.0-10.0

When carrying out wet granulation using a 5% starch paste. The use of 5% starch paste as a humidifier gives the whole mass uniformity and good wettability. For the good solubility of the tablets, a liquid starch paste is necessary, since a thick starch paste increases the rigidity of the entire mass, which reduces the solubility of the tablets. The method of production of such tablets is greatly simplified by the use of one component as a filler and moisturizer. The production of such tablets is inexpensive due to the use of such a component as starch. The use of magnesium stearate as a moving substance, aerosil and talc as dusting substances allows to achieve good fluidity of the granulate, which ensures the normal tabletting process. The introduction of hydroxypropyl cellulose with a low degree of substitution and / or microcrystalline cellulose provides the necessary tablet strength and removes the effect of "capping"; reduces production losses.

Use the sodium salt of 2-methyl-6-nitro-1,2,4-triazolo [5,1-s] -1,2,4-triazin-7-one, a dihydrate, the particle size of which is in the range of 0.1- 0.6 mm.

As a perforated device, a perforated plate is used.

Primary wiping is carried out through a perforated plate with 1.5 mm holes.

For secondary wiping, use a perforated plate with 0.5 mm holes.

Through wet granulation, uniform particle size of the mass is achieved.

The granulate is dried at a temperature of 40-45 ° C to a residual moisture content of not more than 0.5%. Use obtained by the claimed method granulate with higher humidity is impractical, as this leads to a deterioration in flowability (see figure 1).

As can be seen from the graph, the best performance is obtained with a residual moisture content of 0.5%.

Secondary wiping through a perforated plate, the hole size of which is 0.5 mm, allows to obtain granules with improved flowability (see figure 2).

The maximum flowability of particles with a size of from 0.5 to 0.63 mm was determined. Dusting the granules with talcum powder and aerosil prevents them from sticking together.

The granulate obtained is tabletted on a rotary tablet press with a diameter of 9 mm on biconcave punches at 60 MPa to obtain tablet cores with an average weight of 0.340 g. The tablet cores enter a deduster. Then, the tablet cores enter the coater (film coating machine) for applying a 6% hypromellose solution. Coating in an amount of 5% by weight of the tablet core was sufficient to protect the tablet core from the external environment. Received tablets with an average weight of 0.330 g, coated with a protective sheath.

Technical solutions that coincide with the totality of the essential features of the claimed invention have not been identified, which allows us to conclude that the claimed invention meets such a patentability condition as “novelty”.

The claimed essential features that predetermine the receipt of the specified technical result, do not explicitly follow from the prior art, which allows us to conclude that the claimed invention meets such a patentability condition as "inventive step".

The patentability condition “industrial applicability” is confirmed by the example of a specific application.

An example of a specific implementation method:

The components were used in the following ratio: 2-methyl-6-nitro-1,2,4-triazolo [5.1-s] -1,2,4-triazin-7-one sodium salt, dihydrate in the amount of 1200-1800 g, as a filler - potato starch in an amount of 10-400 g, as a baking powder - microcrystalline cellulose in an amount of 10-400 g; as antifriction substances - talc in the amount of 4-50 g, and aerosil - 4-60 g; as a lubricating component - magnesium stearate in an amount of 4-60 g; as a substance for the formation of a protective shell - hypromellose in an amount of 20-200 g.

680 g of 2-methyl-6-nitro-1,2,4-triazolo [5.1-s] -1,2,4-triazin-7-one sodium dihydrate, 82 g of starch are loaded into a high-speed mixer-granulator and 150 g of microcrystalline cellulose and moisturize with 5% starch paste. The moistened mass is passed through a granulator with a hole diameter of 1.5 mm, the granulate is dried in a dryer at a temperature of 40-45 degrees to a residual moisture content of not more than 0.5-1.0%. The obtained granulate is passed through a granulator with a hole diameter of 0.5 mm. The granulate is loaded into a dusting machine and dusted with a mixture of talc (5.5 g), aerosil (14 g) and magnesium stearate (13.5 g). The granulate obtained is tabletted on a rotary tablet press with a diameter of 9 mm on biconcave punches at 60 MPa to obtain tablet cores with an average mass of 0.300 g. The tablet cores enter a deduster. Then, the tablet cores enter the coater (film coating machine) for applying a 6% hypromellose solution. Get tablets with an average weight of 0.315 g, coated with a protective coating, which are placed in the package.

Thanks to the inventive method for producing an antiviral drug in tablets, a drug is created that has a pronounced virus-inhibiting effect, improved organoleptic properties and pharmacological safety.

All components that make up the antiviral agent are approved for use in medical practice and are produced in accordance with the requirements of the State Pharmacopoeia.

The inventive compositions of the antiviral agent and the method for its production expand the arsenal of drugs with a wide spectrum of action, increase the stability of the drug and optimize the process of its preparation. The drug is used to treat diseases caused by bird flu viruses, Rift viruses, parainfluenza virus, Aujeszky's disease virus, Western equine encephalomyelitis, Venezuelan equine encephalomyelitis, and respiratory syncytial virus.

To confirm the achievement of the claimed technical result in the implementation of the invention were used compositions with the contents of the ingredients in the claimed range of values of quantitative characteristics, wt.%, In particular:

Composition No. 1:

Sodium salt of 2-methyl-6-nitro-1,2,4-triazolo [5.1-s] -1,2,4-triazin-7-one dihydrate 76.0 Potato starch 4.0 Low Substitution Hydroxypropyl Cellulose 8.0 Aerosil 2.0 Magnesium stearate 2.0 Talc 2.0 Hypromellose 6.0

Composition No. 2:

Sodium salt of 2-methyl-6-nitro-1,2,4-triazolo [5.1-s] -1,2,4-triazin-7-one dihydrate 71.5 lactose 10 microcrystalline cellulose 12 Aerosil 3.0 Magnesium stearate 0.5 Talc 1,0 Hypromellose 2.0

The implementation of the invention is confirmed by examples of specific performance.

Example 1. The synthesis scheme of the sodium salt of 2-methyl-6-nitro-1,2,4-triazolo [5,1-s] -1,2,4-triazin-7-one, the dihydrate is as follows: to a solution cooled to 0 ° C, containing 13 g (0.1 mol) of 5-amino-3-methylthio-1,2,4-triazole in 16 ml of nitric acid and 100 ml of water, add a solution containing 8 g of sodium nitrite in 50 ml of water for 15 minutes, incubated for 10 minutes and a chilled mixture containing 11 ml (0.1 mol) of ethyl nitroacetate in 150 ml of a 2 M sodium carbonate solution was added portionwise. It is stirred for 2 hours at 0-5 ° C, the precipitate formed is filtered off, then it is successively crystallized from a solution of 20% acetic acid, water and dried in air. Yield: 21.2 g (74%).

Example 2

The study of the antiviral effectiveness of the proposed drug based on structural analogues of purine against the pathogen of influenza A (H5N1)

We used the influenza virus, strain A (H5N1), isolated from biological samples delivered from the outbreak (more than 460 thousand chickens died and were destroyed).

Used transplantable culture of pig kidney cells - SPEV and dog kidneys - MDCK. A semisynthetic medium (PS-4) on Hanks solution containing 7.5% and 2% cattle serum, respectively, was used as the growth and maintenance media.

Used compounds No. 1 and 2

Control preparations: Oseltamivir (Tamiflu) - manufactured by Hoffmann - La Roche Ltd, Switzerland; aribidol - manufactured by ZAO Masterlek, Russia, remantadine - manufactured by OOO ROZFARM, Russia.

The antiviral efficacy of in vitro preparations was evaluated by the following indicators:

- a decrease in the level of virus accumulation under the influence of the drug (Δ, Ig);

- coefficient of inhibition (Ki,%);

- suppression of the cytotoxic activity of the virus (%).

The criteria for the effectiveness of the drugs are: a decrease in the infectious titer of the pathogen by more than 1.8 Ig, Ki more than 98.3%, respectively.

The main criteria for evaluating in vivo efficacy were:

- indicators of protection of laboratory animals from death;

- the average life time of animals in the group (see, for example, Methodological approaches to the search for antiviral drugs, their testing and evaluation / II. A. Lagushkin, N.I. Mitin, V.A. Starovoitova, etc. // Viral inhibitors and the mechanism of their action. - Riga, 1977. - S.138-149).

Statistical processing of the obtained results was carried out by generally accepted methods (see, for example, Lakin GF Biometrics. - M.: Higher School. - 1990).

As an object of study, 9-10-day-old developing chicken embryos (RCEs), white mice weighing 12-14 g and chickens 6-8-week old were used.

Evaluation of the effectiveness of various formulations No. 1 and 2 in the claimed range of quantitative characteristics of the claimed agent in relation to influenza virus, strain A (H5N1), was carried out in vitro in an MDSC cell culture. The allantoic fluid of infected RCEs with biological activity of 5.5 lg CPD / ml was used as an infectious preparation. The multiplicity of infection was 0.01 CPD / cell. Virus adsorption was carried out for 60 min at a temperature of plus (37.0 ± 0.5) ° C. After the adsorption of the virus, the monolayer was washed three times with a PS-4 nutrient medium in a Hanks solution containing 2% cattle serum and 100 IU / ml of penicillin and streptomycin, then fresh medium containing the preparations in the studied concentrations was introduced and incubated for 3 days at a temperature of plus (37.0 ± 0.5) ° С. Throughout the observation period, the cytopathic effect caused by the virus in the cell culture was visually taken into account using a light microscope (objective × 10, eyepiece × 15), and the level of specific hemagglutinin was also determined. Tamiflu, remantadine and arbidol were used as reference preparations. At least 10 tubes with a monolayer of two-day cell culture were used for each dose of the drug.

The results of the study of the suppression of the reproduction of influenza virus, strain A (H5N1) in dynamics, are presented in tables 1 and 2, indicate that the claimed tool (composition No. 1) in the MDCK cell culture in a concentration corresponding to BMD (128 μg / ml), completely inhibits the cytopathic activity of influenza virus, strain A (H5N1), when the drug is introduced 1 hour after infection of the cell monolayer. At concentrations of 64 μg / ml and 32 μg / ml, the product suppresses 90% of the cytopathic activity of the virus.

As can be seen from Table 1, the lowest antiviral efficacy was revealed by arbidol. In the maximum tolerated concentration, the inhibition coefficient was 30%.

Oseltamivir in the maximum tolerated concentration completely suppresses the cytopathic activity of the avian influenza virus, and the effectiveness of remantadine was 87%.

As a result of studying the ability of the claimed drug to suppress the formation of specific hemagglutinin, it was revealed that it suppresses the reproduction of influenza virus in the maximum tolerated concentration by 93.8%, at a concentration of 64 μg / ml - by 89.6%, at a dose of 32 μg / ml - by 87 ,5%. The most effective suppresses Tamiflu virus reproduction at the maximum tolerated concentration.

As can be seen from Table 2, the claimed agent in high concentrations effectively inhibits the reproduction of influenza virus, strain A (H5N1), in the MDCK cell culture.

The results of a study of the suppression of reproduction of influenza virus, strain A (H5N1), are presented in table 2, indicate that the agent in the culture of MDCK cells at a concentration corresponding to 1/16 IPC (8 μg / ml) inhibits the cytopathic activity of influenza virus, strain A (H5N1).

Arbidol showed the lowest antiviral efficacy. In the maximum tolerated concentration, the inhibition coefficient was 50%. Oseltamivir and remantadine in the maximum tolerated concentration completely suppress the cytopathic activity of avian influenza virus.

As a result of studying the ability of the claimed drug to suppress the formation of specific hemagglutinin, it was found that the drug in 1/16 of the maximum tolerated concentration suppresses the reproduction of the influenza virus completely, compounds No. 1 and 2. Tamiflu and remantadine effectively suppress the reproduction of the virus in the maximum tolerated concentration.

Thus, the evaluation of the antiviral efficacy of various formulations of the claimed agent showed their reproducibility in the process of suppressing reproduction of the influenza virus.

Example 3

The study of the effectiveness of the claimed funds against influenza virus strain A (H5N1) in vivo

Evaluation of the effectiveness of the proposed drug against influenza virus, strain A (H5N1), was carried out in vivo using white mice weighing 12-15 g. Allantoic fluid of infected RCEs with biological activity of 5.5 lg CPD ₅₀ / ml, 8 was used as an infectious preparation. , 3 lg ELD ₅₀ / ml; 6.3 lg LD ₅₀ / ml.

The preparations were administered orally to white mice according to the prophylactic and therapeutic regimens, as well as according to the emergency prophylaxis regimen.

The results of a study of the preventive efficacy of the Agent in relation to the experimental form of influenza in white mice intranasally infected with the influenza virus are presented in Table 4.

The protection against death when using this compound at a dose of 100 mg / kg of the animal's weight averaged 35.0%, the increase in average life time (LSS) was 2.6 days.

The use of the Means according to the prophylactic scheme at a dose of 50 mg / kg protected an average of 31.7% from the death of infected animals; at a dose of 10 mg / kg - 33.3%; 1 mg / kg - 45.0%. The increase in the SVZ indicator averaged, according to the results of the experiments, by 2.4 days, 2.6 days and 2.8 days, respectively.

To select the most effective regimen for the use of the drug, the drug was administered orally to animals 2 days and a day before infection. The results of the study are presented in Table 5.

When using the Agent twice in 48 hours and 24 hours before infection, the drug provided more effective protection to animals at doses of 1 and 10 mg / kg. Protection against death amounted to 40.0%, and an increase in the indicator of life expectancy - 3 days. The antiviral effectiveness of the drug is comparable with the same indicator for arbidol. It should be emphasized that the dose of the drug was significantly lower than the reference drug arbidol, and a day more CSF in the group.

When using the Agent one day and 1 hour before infection, the protective effectiveness indicator of the Agent decreased by 10%.

Arbidol and Tamiflu were used as reference preparations. Arbidol at a dose of 30 mg / kg (corresponding to a prophylactic dose for humans - 200 mg / day) protected from death on average 38% of infected white mice. The increase in average life time averaged 2.2 days. At a dose of 60 mg / kg (corresponding to an equally effective human dose - 400 mg / day), the protective efficacy averaged 45%, an increase in the average life time of 2.6 days.

The protective efficacy of Tamiflu 10 mg / kg was on average 51% at a dose of 13 mg / kg (corresponds to _^1/2 human dose - 75 mg / day) - 62% at a dose of 50 mg / kg - 86%. At the same time, an extension of the average life time by 3.4 was observed; 4.3 and 4.7 days, respectively.

Therefore, the greatest antiviral effect of the tool was shown when applied in the dose range from 1 to 10 mg / kg of the animal’s weight according to prophylactic regimens 5 days before infection, daily and 2 days before infection daily.

When using the Means according to the emergency prevention scheme at a dose of 50 mg / kg of the mass, the drug protected from death on average 42.5% of infected animals, at a dose of 100 mg / kg - 47.5%. At the same time, an increase in the CSW was observed by 2.7 days and 3.0 days, respectively.

Arbidol at a dose of 135 mg / kg, when used according to the emergency prevention scheme, protected 30% of white mice infected with the influenza virus from death. The increase in SVZh averaged 1.7 days.

Example 4

Tests of antiviral activity of the claimed funds against the virus of "swine" flu

Study of protective activity. Means against the swine flu virus were carried out on the basis of the research institute of influenza. The objectives of the study were to reproduce the infectious process caused by this virus in white mice and to test the antiviral activity of the drug in comparison with the antiviral drug Remantadin.

In the work, aliquots of the Means were diluted in Cell Culture Medium Needle MEM. The necessary dilutions were prepared from the resulting solution on MEM medium for animal experiments. Remantadine (1- (1-adamantyl) aminoethyl hydrochloride, Aldrich Chem. Co.) was used as a reference preparation. Influenza virus A / California / 07/09 (H1N1) from the collection of viral strains of the research institute of influenza was used as a model virus. The virus was passaged in the allantoic cavity of 10-12 day old chicken embryos for 48 hours at 36 ° C.

The studied preparations were administered intraperitoneally to animals in a volume of 0.2 ml in the following doses: Agent 200 and 70 mg / kg, Remantadine - 50 mg / kg. The drugs were administered according to the treatment-and-prophylactic regimen: 24 hours and 1 hour before infection and 24, 48 and 72 hours after infection. As a placebo, a control group of animals was injected with physiological phosphate buffer. Intact animals that were kept under the same conditions as the experimental groups were used as a negative control. Viruses were administered to animals intranasally under mild ether anesthesia at a dose of 1 LD ₅₀ . 15 mice were taken in each observation group. On day 3 after infection, 5 animals from each group were sacrificed, lungs were opened and isolated. Lungs were frozen and stored at -20 ° C until appropriate experiments were performed.

The remaining animals were monitored for 14 days, i.e. the period during which animal mortality is observed in experimental flu. Daily recorded mortality of animals in the control and experimental groups. Based on the obtained mortality rates in each group, the percentage of mortality was calculated (M, the ratio of the number of animals killed during 14 days to the total number of infected animals in the group), the protection index (IP, the ratio of the difference in percent of mortality in the control and experimental groups to the percentage of mortality in the control group ) and the virus in white mice, and testing the antiviral activity of the Means in comparison with the antiviral drug Remantadin.

Thus, the developed antiviral agent in the form of tablets has a pronounced virus-inhibiting effect, pharmacological safety and improved organoleptic properties.

Table 1 The results of evaluating the effectiveness of means for suppressing the cytopathic activity of influenza virus, strain A (H5N1), in an MDCK cell culture 48 hours after infection (n = 3) A drug Dose, mcg / ml The frequency of detection of the CPP, X CPD rejection ratio%, X The inventive tool composition No. 1 128.0 0/30 one hundred 64.0 3/30 90 32,0 3/30 90 Arbidol 25.0 21/30 thirty Tamiflu 100.0 0/30 one hundred Remantadine 50,0 4/30 87 Control without drug one hundred -

table 2 The results of evaluating the effectiveness of means for suppressing the formation of specific hemagglutinin of influenza virus, strain A (H5N1), in an MDCK cell culture 48 hours after infection (n = 3) A drug Dose, mcg / ml Hemagglutinin level, GA, X ± σ _x GA suppression coefficient,%. X ± a _x The inventive tool composition No. 1 128.0 8.0 ± 0.0 93.8 ± 0.0 64.0 13.2 ± 2.5 89.5 ± 2.1 32,0 16.0 ± 0.0 87.5 ± 0.0 Arbidol 25.0 53.2 ± 10.6 58.2 ± 8.3 Tamiflu 100.0 absent 100.0 ± 0.0 Remantadine 50,0 1.3 ± 0.7 98.9 ± 0.5 Control (without drug) 128 ± 0,0 -

Table 3 Evaluation of the antiviral efficacy of various formulations of the Agent against influenza virus, strain A (H5N1) in MDCK cell culture (by cytopathic activity) (n = 3) A drug Dose of the drug. mcg / ml The frequency of detection of the CPP, X The coefficient of suppression of the CPU%. X ± σ _x The inventive Tool, Composition No. 1 8.0 6/30 80 4.0 9/30 70 2.0 12/30 60 1,0 15/30 fifty 0.5 18/60 40 The inventive Tool, Composition No. 2 8.0 3/30 90 4.0 18/30 80 2.0 12/30 60 1,0 15/30 fifty 0.5 15/30 fifty Arbidol 25 15/30 fifty Tamiflu one hundred 0/30 one hundred Remantadine fifty 0/30 one hundred I Control - 30/30 -

Table 4 Study of the prophylactic efficacy of the Agent in relation to the experimental form of influenza in white mice infected intranasally with the influenza virus, strain A (H5N1) A drug Scheme of drug administration Dose, mg / kg Animal death rate Protective efficiency from death,% The average life time of animals in the group, days Increase in average life expectancy, Δ, days The inventive tool. Composition No. 2 - 120 hours, - 96 hours; - 72 hours; - 48 hours; - 24 hours; - 1 h one hundred 10/20 fifty 11,2 2.7 fifty 14/20 thirty 10.8 2,3 10 13/20 35 10.8 2,3 one 10/20 fifty 11,4 2.9 Tamiflu fifty 4/20 80 10.0 4.0 13 8/20 60 12.5 4.0 10 10/20 fifty 11.5 3.0 Arbidol Dose Control - 24 hours, - 1 hour 60 11/20 45 11.1 2.62.5 thirty 12/20 40 11.0 Dose control - - 20/20 - 8.5 - Herd control - - 0/20 - 14.0 - Note 1 The infectious dose was 10 LD ₅₀

Table 5 Study of the prophylactic effectiveness of the Agent in relation to the experimental form of influenza in white mice infected intranasally with the influenza virus, strain A (H5N1) A drug Scheme of drug administration Dose, mg / kg Animal death rate Protective efficiency from death,% The average life time of animals in the group, days Increase in average life expectancy, Δ, days The inventive tool composition No. 2 - 24 hours one hundred 15/20 25 11.0 2,3 fifty 14/20 thirty 11,2 2.5 10 14/20 thirty 11.3 2.6 one 14/20 thirty 11,4 2.7 Tamiflu - 120 hours, - 96 hours, - 78 hours, - 48 hours, - 24 hours, - 1 hour fifty 2/20 90 13.6 4.9 13 7/20 65 13,2 4,5 10 10/20 fifty 12.5 3.8 Arbidol - 24 hours, - 1 hour 60 11/20 45 11.5 2,8 thirty 12/20 40 11.0 2,3 Dose control - - 20/20 - 8.7 - Herd control 0/20 14.0

Table 6 The indicators of the pathological process in influenza pneumonia of white mice caused by the influenza virus A / California / 7/09 (H1N1), in the conditions of use of the claimed Means The drug, mg / kg average life expectancy Mortality% Protection Index,% Increase in life expectancy The inventive tool composition No. 1 200 mg / kg 13.7 twenty 60 3,1 The inventive tool, composition No. 1, 70 mg / kg 12.8 thirty 40 2.2 Remantadine 11.5 40 twenty 0.9 Virus control 10.6 fifty - 0

Claims (10)

1. An antiviral agent in tablets containing the active substance, physiologically acceptable auxiliary additives, namely a filler, a disintegrant, an antifriction substance, a lubricant and a protective coating, characterized in that 2-methyl-6-nitro sodium salt is used as the active substance 1,2,4-triazolo [5,1-c] -1,2,4-triazin-7-one, dihydrate, as filler, potato starch and / or lactose, as a disintegrant, hydroxypropyl cellulose with a low degree of substitution and / or microcrystalline cellulose; as antifriction substances - talc and aerosil; as a lubricating component - magnesium stearate; as a substance for the formation of a protective shell - hypromellose, in the following ratio of components, wt.%:
Sodium salt of 2-methyl-6-nitro-1,2,4-triazolo [5.1-s] -1,2,4-triazin-7-one dihydrate 60.0-90.0 Potato starch and / or lactose 1,0-20,0 Low Substitution Hydroxypropyl Cellulose and / or microcrystalline cellulose 0.5-20.0 Aerosil 0.2-3.0 Magnesium stearate 0.2-3.0 Talc 0.2-2.5 Hypromellose 1.0-10.0 2. The antiviral agent in tablets according to claim 1, characterized in that the granule size of the agent is in the range from 0.5 to 0.63 mm. 3. The antiviral agent in tablets according to claim 1, characterized in that the tablet is made weighing from 0.300 to 0.330 g. 4. The antiviral agent in tablets according to claim 1, characterized in that it has a virus inhibitory activity against avian influenza virus, Rift Valley fever viruses, parainfluenza, Aujeszky's disease virus, equine horse encephalomyelitis, equine venezuelan encephalomyelitis, and respiratory syncytial virus. 5. A method of obtaining an antiviral agent in tablets, carried out using the active substance, physiologically acceptable auxiliary additives: filler, disintegrant, anti-friction substance, lubricant and protective coating and comprising moisturizing, subsequent wet granulation, drying, dry granulation, dusting, tabletting and application film coating, characterized in that when mixing the active substance, filler, baking powder, wet granulation of the mass is carried out, after which is the primary wiping moistened mass through a perforated device, dried, and the drying is carried out to a residual moisture content of not more than 0.5%; then secondary rubbing is carried out through a perforated device, after which the granulate obtained is dusted with a mixture of talc, aerosil and magnesium stearate, then tableted and coated with a protective coating, while the sodium salt of 2-methyl-6-nitro-1,2,4 is used as the active substance triazolo [5,1-c] -1,2,4-triazin-7-one, dihydrate, potato starch and / or lactose as a filler, low substitution hydroxypropyl cellulose and / or microcrystalline cellulose as a disintegrant; as antifriction substances - talc and aerosil; as a lubricating component - magnesium stearate; as a substance for the formation of a protective shell - hypromellose, in the following ratio of components, wt.%:
Sodium salt of 2-methyl-6-nitro-1,2,4-triazolo [5.1-c] -1,2,4-triazin-7-one dihydrate 60.0-90.0 Potato starch and / or lactose 1,0-20,0 Low Hydroxypropyl Cellulose substitution and / or microcrystalline cellulose 0.5-20.0 Aerosil 0.2-3.0 Magnesium stearate 0.2-3.0 Talc 0.2-2.5 Hypromellose 1.0-10.0 6. The production method according to claim 5, characterized in that when carrying out wet granulation using 5% starch paste. 7. The production method according to claim 5, characterized in that the sodium salt of 2-methyl-6-nitro-1,2,4-triazolo [5,1-s] -1,2,4-triazin-7-one is used dihydrate, the particle size of which is in the range of 0.1-0.6 mm 8. The production method according to claim 5, characterized in that a perforated plate is used as the perforated device. 9. The production method according to claim 7, characterized in that the primary wiping is carried out through a perforated plate with 1.5 mm holes. 10. The production method according to claim 7, characterized in that when the secondary wiping use a perforated plate with holes of 0.5 mm

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