RU2429875C2 - Pharmaceutical composition of antiviral preparation and method for prevention and treatment of viral diseases by this preparation - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention represents a pharmaceutical composition for treating viral diseases, containing dipeptide, differing by the fact that dipeptide is an aqueous solution of dipeptide of formula (I) Gly-Pro in concentration 0.9-1.1 %; besides, Nipagin is used as a preservative; the ingredients of the composition are taken in certain proportions, g/l.

EFFECT: extended range of the antiviral drugs, more efficient prevention and treatment of viral infection.

3 cl, 5 ex, 5 tbl

Description

The invention relates to the field of pharmacology and medicine, namely to the development of a pharmaceutical composition and method of using an antiviral preparation based on the dipeptide of the formula (I) Gly-Pro having an antiviral effect.

The relevance of the development of new antiviral drugs is determined by the wide spread of viral diseases such as influenza, herpes, cytomegalovirus infections, hepatitis, etc., and the lack of drugs for their treatment. In recent years, there has been a population decline in immunity associated with a deterioration in the human environment, which also contributes to an increase in viral diseases.

A known method for the prevention and treatment of viral diseases, including the use of a medicinal product [Ridostin, Cycloferon, Dalargin, Grippferon, Arbidol-Lance, Acyclovir-Acre]. However, this known method is not in all cases effective.

A known pharmaceutical composition for treating viral diseases, comprising 0.004-0.040 g of 2-benzylbenzimidazole hydrochloride hydrochloride (dibazole), 0.025-0.100 g of ascorbic acid and 0.0005-0.005 g of L-glutamyl-L-tryptophan monosodium salt dipeptide (thymogen) as immunomodulator, which can be considered as a prototype [RF patent No. 2165254]. In this composition, dibazole acts as an inducer of endogenous interferon, the dipeptide is an immunomodulator, ascorbic acid is an antioxidant. The prototype provides a pharmaceutical composition that is assembled in a single dosage form, which can be packaged in gelatin capsules or available in the form of tablets or coated tablets. The disadvantage of this known composition is the complexity of its production, requires special measures to protect the dipeptide in the gastrointestinal tract from proteolysis.

The specified technical result is achieved by the use of the dipeptide of the formula (I) Gly-Pro as an antiviral drug in aqueous solution. Similar peptides related to glyprolins are stable in the gastrointestinal tract and at the same time, as the examples show, have a specific antiviral effect. A pharmaceutical composition containing a dipeptide of formula (I) Gly-Pro is easy to manufacture, which greatly reduces the cost of the drug.

A known method for the prevention and treatment of viral diseases through the use of the Selank heptapeptide of the formula Thr-Lys-Pro-Arg-Pro-Gly-Pro by intranasal administration of its 0.12-0.18% solution in a daily dose of 550-650 μg for 10- 18 days [RF patent No. 2337703, 11/10/2008, bull. No. 31, Andreeva L.A., Ershov F.I., Zozulya A.A., Mezentseva M.V., Myasoedov N.F., Seredenin S.B., Surgucheva I.M., Uchakina O.N. , Uchakin PN, Means for the prevention and treatment of viral diseases and a method for the prevention and treatment of viral diseases].

This method can be considered the prototype of the invention.

The technical result achieved by the implementation of the present invention is to expand the range of antiviral drugs, increase the effectiveness of the prevention and treatment of viral diseases, reduce the cost of the drug by significantly simplifying the synthesis of the proposed dipeptide of formula (I) Gly-Pro instead of the Selank heptapeptide.

The specified technical result is achieved by the fact that the dipeptide of the formula (I) Gly-Pro is used as a means of prevention and treatment of viral diseases of viral etiology (influenza, herpes, cytomegalovirus infections) in the following pharmaceutical composition.

A pharmaceutical composition for treating viral diseases comprising an aqueous solution of the dipeptide of formula (I) Gly-Pro, characterized in that it contains a dipeptide of formula (I) Gly-Pro in a concentration of 0.9-1.5%, additionally nipagin as a preservative in the following ratio of components, g / l:

dipeptide of the formula (I) 9.0-15.0 nipagin 0.9-1.1 distilled water up to 1 liter

This technical result is achieved due to the method for the prevention and treatment of viral diseases, involving the use of a drug, where a dipeptide of the formula (I) Gly-Pro is used as a drug by intranasal administration of its 0.9-1.5% solution in a daily dose 12-20 mg / day for 2-3 days, namely:

a method for the prevention of viral diseases, including the introduction of a pharmaceutical composition of 2 drops in each nasal passage 6 times a day for 2 days;

a method for the treatment of viral diseases, including the introduction of a pharmaceutical composition of 3 drops in each nasal passage 7 times a day for 3 days.

Our own studies have found that the drug based on the dipeptide of formula (I) Gly-Pro is able to suppress the reproduction of influenza viruses, herpes simplex virus (HSV) types 1 and 2, cytomegalovirus infections and can be used for the prevention and treatment of viral diseases caused by viruses, without causing adverse side effects.

The following are examples illustrating the invention.

Example 1. The study of the effect of the route of administration of the drug based on the dipeptide of formula (I) Gly-Pro on antiviral activity against encephalomyocarditis virus (VEMK)

Studies were carried out on white outbred mice weighing 14-16 grams of 5 mice for each method of administration. Animals were infected with encephalomyocarditis virus (VEMK).

A preparation based on the dipeptide of formula (I) Gly-Pro in concentrations of 1%; 0.5%; 0.2%; 0.1% supplemented with 0.1% nipagin as a preservative was administered to animals intraperitoneally, intranasally, orally at 0.2 ml / mouse at the time of infection.

The control group consisted of mice infected with the flu and given a placebo. The observation period is 15 days.

The antiviral effect of the drug was evaluated by the survival of mice (% survival,% protection, average harmonic value of the lifetime).

Evaluation of the activity of the drug was carried out, given:

1) the average harmonic value of the lifetime for each group of mice, including information on all individuals, calculated by the formula:

1 / τ = (X ₁ / t ₁ + X ₂ / t ₂ + ... + X _a / t _a ): n, where t is the day of death of animals, X is the number of dead animals on day t, and is the day of observation; n is the number of animals in the group (5 mice);

2) the ratio of protection (KZ) - the ratio of the decrease in the number of sick and dead animals in the experimental group compared with the control, calculated by the formula:

KZ =% dead animals in the control group /% dead animals in the experimental group;

3) the drug effectiveness index (IE), calculated by the formula:

IE = [(KZ-1) / KZ] × 100%.

Table 1 The antiviral activity of a preparation based on the dipeptide of formula (I) Gly-Pro with different methods of administration for encephalomyocarditis virus No. p.p. Route of administration Drug concentration % survival % protection τ * one orally 0.5 twenty twenty 7.63 2 intranasally 0.5 40 40 8.33 3 intraperitoneally 0.5 twenty twenty 7.14 four the control 100 LD50 0 0 5.35 Note: τ * is the average harmonic value of the life time of animals.

Studies of the antiviral activity of the preparation based on the dipeptide of formula (I) Gly-Pro in relation to VEMC showed that the preparation based on the dipeptide of formula (I) Gly-Pro helps to protect 20-40% of mice, depending on the method of administration. The greatest protection was observed with the intranasal route of administration of the drug.

Example 2. The study of the method of administration of the drug based on the dipeptide of the formula (I) Gly-Pro for antiviral activity against influenza virus A / Aichi / 1/68

Studies were carried out on white outbred mice weighing 14-16 grams of 5 mice for each method of administration. Animals were infected with influenza virus A / Aichi / 1/68. A preparation based on the dipeptide of formula (I) Gly-Pro at a concentration of 0.5% with the addition of 0.1% nipagin was administered as a preservative to animals intraperitoneally, intranasally, orally at 0.3 ml / mouse at the time of infection. The control group consisted of 10 mice infected with the flu and given a placebo. The observation period is 15 days. The antiviral effect of the drug was evaluated by the survival rate of mice (protection ratio (KZ), drug efficacy index (IE), τ * is the average harmonic value of the animal's lifespan).

table 2 The antiviral activity of the drug based on the dipeptide of formula (I) Gly-Pro with different methods of administration against influenza virus A / Aichi / 1/68 No. p.p. Route of administration Drug concentration Multiplicity of protection (KZ) Drug Performance Index (IE) one orally 0.5 1.25 twenty 2 intranasally 0.5 1,67 60 3 intraperitoneally 0.5 1.25 40 four virus control 100 LD50 1,0 0

A study of the antiviral activity of a preparation based on the dipeptide of formula (I) Gly-Pro against influenza virus A / Aichi / 1/68 showed that the preparation based on the dipeptide of formula (I) Gly-Pro helps to protect 20-40% of mice, depending on the method introduction. The greatest protection was observed with the intranasal route of administration.

Example 3. The study of the effect of the dose of the administered drug based on the dipeptide of the formula (I) Gly-Pro on the antiviral activity against mouse encephalomyocarditis virus (VEMK)

Studies were conducted on mice weighing 14-16 grams - 5 mice per point. Animals were infected with encephalomyocarditis virus (VEMK) at a dose of 100 LD50.

A preparation based on the dipeptide of formula (I) Gly-Pro in concentrations of 1%; 0.5%; 0.2%; 0.1% supplemented with 0.1% nipagin as a preservative was administered to animals intranasally at 0.2 ml / mouse at the time of infection. The control group consisted of mice infected with VEMK and receiving a placebo. The observation period is 15 days. The antiviral effect of the drug was evaluated by the survival of mice (% survival,% protection, average harmonic value of the lifetime) according to the formulas above (see example 2). The data are shown in table 3.

Table 3 The dependence of the antiviral activity against VEMK virus on the dose of the drug based on the dipeptide of the formula (I) Gly-Pro No. p.p. The administered dose of the drug,% % survival % protection τ * one 1,0 twenty twenty 6.76 2 0.5 40 40 8.33 3 0.2 twenty twenty 8.06 four 0.1 40 40 9.80 the control 0 0 0 5.35

As can be seen from the above data, the best result with respect to the encephalomyocarditis virus is achieved by intranasally administering a 0.09-0.15% solution of the drug with the addition of 0.1% nipagin as a preservative, which corresponds to the administration of about 12-14 mg / kg per day .

Example 4. The study of the effect of the dose of the injected drug based on the dipeptide of formula (I) Gly-Pro on antiviral activity against influenza virus A / Aichi / 1/68

Studies were carried out on white outbred mice weighing 14-16 grams - 5 mice for each method of administration. Animals were infected with influenza virus A / Aichi / 1/68.

A preparation based on the dipeptide of formula (I) Gly-Pro in concentrations of 1%; 0.5%; 0.2%; 0.1% was administered to animals intranasally at 0.3 ml / mouse at the time of infection.

The control group consisted of 10 mice infected with the influenza A / Aichi / 1/68 virus and treated with placebo. The observation period is 15 days. The antiviral effect of the drug was evaluated by the survival of mice (protection ratio (KZ), drug efficacy index (IE)). The data obtained are shown in table 4.

Table 4 The dependence of antiviral activity on the dose of the drug No. p.p. The administered dose of the drug (% solution of the drug) Multiplicity of protection (KZ) Drug Performance Index (IE) one 1,0 1.25 twenty 2 0.5 1,67 40 3 0.2 1.25 twenty four 0.1 1,67 40 the control 0 1,0 0

As can be seen from the above data, the best result with respect to the A / Aichi / 1/68 influenza virus is achieved by intranasally administering 0.3 ml of a 0.09-0.15% solution of the drug containing 0.1% nipagin as a preservative, which corresponds to the introduction of 18-20 mg / kg per day.

Thus, the data shown in tables 3-4 show that doses of 12-20 mg / kg per day with the intranasal route of administration provide antiviral protection for animals.

Example 5. The study of the antiviral activity of the drug based on the dipeptide of the formula (I) Gly-Pro with different schemes of administration

We studied the effectiveness of the drug in infections caused by the mouse encephalomyocarditis virus (VEMC), with the introduction of the following schemes:

prevention -

1) 24 hours before infection;

2) 2 consecutive days 24 and 48 hours before infection;

treatment -

1) 24 hours after infection;

2) 24 and 48 hours after infection;

3) 3 times in a row after 24, 48 and 72 hours after infection.

The drug was administered, based on previous experiments, intranasally.

Infection was performed with mouse encephalomyocarditis virus (VEMK) at a dose of 100 LD50.

The results of the study of the antiviral activity of the drug for different schemes of administration against influenza virus in the model of experimental infection caused by VEMK in mice are presented in table 5. It is shown that the drug protected animals from the action of VEMK. There were 3.5 surviving animals with a prophylactic regimen of drug administration compared to the control and 4 times more when using a drug based on the dipeptide of formula (I) Gly-Pro for the treatment of mice.

Table 5 The survival of mice with different patterns of drug administration Scheme of drug administration % survival Preventative
1-time administration 24 hours before infection fifty Preventative
2-time administration 48 and 24 hours before infection 70 Treatment
1-time administration 24 hours after infection 60 Treatment
2-time administration after 24 infections and 48 hours after infection 60 Treatment
3 times 24, 48 and 72 hours after infection 80 The control twenty

The data obtained showed that the greatest antiviral effect against VEMC is achieved with the introduction of the drug before infection once a day for two days, and in the case of treatment after infection with the virus - once a day for three days.

In general, the given examples show the antiviral effect of the drug with the intranasal method of administration to animals at a dose of 12-20 mg / kg per day. For prevention, the drug is administered within two days, in the case of treatment - within three days.

From the data given in tables 3 and 4, it follows that the effectiveness of the drug with the intranasal method of administration to mice using 0.1%; 0.2%; 0.5%; A 1.0% solution of the drug with the addition of 0.1% nipagin as a preservative is approximately the same. However, to create a daily dose of 12-20 mg / kg per day, it is convenient to use a 1% solution of the drug. In this case, a daily dose of 12 mg / day corresponds to 24 drops, i.e. 2 drops in each nasal passage 6 times a day, and a daily dose of 20 mg / day corresponds to 40 drops, i.e. 3 drops in each nasal passage 7 times a day.

Claims (3)

1. A pharmaceutical composition for the treatment of viral diseases, containing a dipeptide, characterized in that the dipeptide is an aqueous solution of the dipeptide of the formula (I) Gly-Pro in a concentration of 0.9-1.1%, additionally nipagin as a preservative in the following ratio of components g / l:
dipeptide of the formula (I) 9.0-11.0 nipagin 0.9-1.1 distilled water up to 1 l 2. A method for the prevention of viral diseases, including the introduction of the pharmaceutical composition according to claim 1, 2 drops in each nasal passage 6 times a day for 2 days. 3. A method of treating viral diseases, including the introduction of the pharmaceutical composition according to claim 1, 3 drops in each nasal passage 7 times a day for 3 days.