WO2007040426A1

WO2007040426A1 - Antiviral agent

Info

Publication number: WO2007040426A1
Application number: PCT/RU2006/000516
Authority: WO
Inventors: Vladimir Nikolaevich Hudoley
Original assignee: Vladimir Nikolaevich Hudoley
Priority date: 2005-10-05
Filing date: 2006-10-04
Publication date: 2007-04-12
Also published as: RU2005130751A

Abstract

The invention relates to medicine, in particular to pharmacology and can be used in the form of an antiviral agent for influenza treatment and prevention. The aim of said invention is to deliver a non-toxic medicinal agent, which is devoid of side effects and exhibits a high antiviral activity. For this purpose, the inventive antiviral agent also contains 3-67 mass % remantadin and 23-97 mass % iodoantipyrine.

Description

ANTIVIRAL AGENT

Technical field

The invention relates to medicine, specifically to pharmacology, and can be used as an antiviral agent for the treatment and prevention of influenza.

BACKGROUND OF THE INVENTION Influenza and acute respiratory viral infections take first place in the frequency and number of cases in the world, and make up 95% of all infectious diseases. Up to 500 million people fall ill every year in the world, 2 million of whom die. In Russia, annually from 27.3 to 41.2 million sick flu and other acute respiratory viral infections are recorded. During epidemics, the level of hospitalization increases by 2-5 times. A particularly high level of hospitalization in young children (under the age of 5 years), and in the elderly (over 65 years). Each influenza epidemic is accompanied by increased mortality. Influenza mortality rates in the world are 0.01-0.2%, and average annual losses reach tens of thousands of people across countries, including mainly children of the first years of life (up to 2 years) and elderly people (over 65 years). Influenza collects the largest victims among the elderly population suffering from chronic diseases (high-risk individuals).

Therefore, it seems very relevant to search for new drugs that have, on the one hand, low toxicity and do not cause side effects, and on the other, have high antiviral activity.

One of the most well-known means for preventing and treating influenza is remantadine, which contains α-methyl (l-adamantyl) -methylamine hydrochloride as an active substance [1]. The prophylactic use of this drug can reduce the incidence of The toxic use of this drug reduces the incidence of influenza by 50%. For maximum effectiveness, you must take this tool throughout the epidemic.

However, it should be noted that remantadine in the conventional dosage form has a number of significant drawbacks that limit its use in high dosages for adults and preschool children: 1) the drug is active only against influenza A virus; 2) when taking the drug, side effects are recorded in relation to the central nervous system, liver and kidney cells, the gastrointestinal tract (most often in the elderly); 3) the rapid formation of resistant strains of the virus. In patients from whom the drug-resistant viruses were isolated, there was no clinical deterioration, but the disease lasted longer. In addition, the rapid removal of the drug from the body requires frequent administration to maintain the necessary therapeutic dose, which in turn leads to undesirable side effects.

Recently, the attention of doctors has been increasingly attracted to the means of non-specific prophylaxis of influenza, acute respiratory viral infections and other infections. Here in the foreground are the interferon inducers. Interferon - a protein produced in the body, has a wide range of antiviral activity, covering, including all known varieties of influenza. One of the most promising drugs in this regard - interferon inducers - is iodantipyrine.

The use of iodantipyrine for the treatment and prevention of tick-borne encephalitis induced in the experiment is known [2]. The use of iodantipyrine for the prevention of influenza and other acute respiratory viral infections has become possible due to the identification of its protective properties against influenza viruses and other acute respiratory viral infections [3]. However, side effects from the gastrointestinal tract and central nervous system observed during treatment iodantipyrine, limit the possibility of its use for children and the elderly.

There are a number of antiviral drugs that include remantadine. For example, a pharmaceutical composition is known that contains remantadine as the active substance and as target additives disaccharide, starch, talc, stearic acid and polyvinylpyrrolidone in the following ratio of components, May. %: remantadine 20-50; disaccharide - 30-70; starch - 5-20; talc - 0.3-3; stearic acid - 0.3-1; polyvinylpyrrolidone - 0.5-3 [4]. Known pharmaceutical composition containing remantadine from 7.0-33.0 may. % and no-shpu from 13.0 - 53.0 May. % [5].

The closest in technical essence and the achieved result to the claimed invention is the connection of remantadine with a copolymer of vinyl alcohol and N-vinylmidosuccinic acid due to the formation of a C-O bond under the name "Polymer" [6]. This agent has several advantages over native remantadine, because it has a more pronounced antiviral effect and has an interferonogenic effect.

However, the known preparation has side effects affecting the change in the acidity of the stomach, and its immunomodulatory properties are insufficient for a pronounced antiviral effect.

SUMMARY OF THE INVENTION An object of the present invention is to provide a non-toxic drug devoid of side effects and having high antiviral activity.

This object is achieved in that the known antiviral agent containing rimantadine additionally contains iodine tipirin in the following ratio, May. %:

Remantadine - 3 - 67

Iodantipyrine - 23 - 97

With this simultaneous use of drugs, the dose of remantadine was reduced, which significantly reduces the formation of resistant strains of the influenza virus and enhances the inhibition of the reproduction of influenza A and B viruses [7]. In addition, the combined use of remantadine and iodantipyrine provides a significant (not less than 4 times) increase in antiviral activity and reduces toxic effects compared with the separate use of these drugs.

The claimed ratio of iodantipirin and remantadine allows you to get a drug with high therapeutic activity that does not cause side effects in patients when it is taken. It was shown for the first time that the studied pharmacological composition in the proposed ratios exerts a pronounced antiviral effect in the treatment, prophylactic, therapeutic and prophylactic regimens of the experimental model of influenza infection. The authors did not find in the literature they analyzed information about the antiviral activity of the proposed pharmaceutical composition with the indicated ratio of the components. Empirically established new properties and possibilities of using the claimed funds. Thus, the proposed solution meets the criteria of the invention of "novelty", "inventive step", "industrial applicability". To confirm these properties, a number of experimental studies were carried out: determining the effectiveness of both individual components and the pharmacological composition itself on a model of experimental influenza infection during treatment and prophylaxis (5- multiple administration of the drug 24 hours and 1 hour before infection with the influenza virus, 24, 48, 72 hours after infection), separate treatment (24, 48, 72 hours after infection) and prophylactic (48, 24 and 1 hour before infection) schemes, determination of the concentration of influenza virus in the lungs of mice on the 1st, 3rd, 5th, 7th day from the moment of infection, the study of the blood serum of experimental mice for the presence of interferon and antihemagglutinins.

The results were processed statistically, by the methods of parametric and nonparametric statistics. The differences were considered significantly significant at P _u <0.05 and P _t <0.05 for quantitative characteristics (Wilcoxon-Mann-Whitney test and Student's test, respectively); at P <0.05 - for qualitative ones (the Fisher angular transformation method) [8].

Experiments with the influenza virus were performed on 495 non-inbred mice weighing 22-25 g; in immunological studies, 160 non-inbred male mice were used, as well as 40 BALB / s mice weighing 18-20 g. Animals were kept on a normal diet in vivarium conditions. The experiments were performed using test viruses obtained from the State Virus Collection of the Museum of Strains of the Institute of Virology of the Russian Academy of Medical Sciences: influenza A virus (Aichi / 2/68), adapted for reproduction in lungs of mice with a titer of 5.0 Ig LD ₅ o.

Studies have shown that the simultaneous use of these drugs provides a more effective inhibition of the reproduction of influenza virus. The following advantages were noted compared with the use of remantadine alone: a) an increase in antiviral activity, b) a decrease in the dose of remantadine makes it possible not only to relieve side symptoms (from the central nervous system, liver and kidney cells, and the gastrointestinal tract), but also dramatically reduce the incidence of rimantadine-resistant strains of type A influenza viruses. It is known that the formation of rimantadine-resistant strains of influenza virus is significantly reduced at low doses of rimantadine. In order to compare the effectiveness of the proposed pharmaceutical composition and its individual components, the therapeutic and prophylactic effect of separately 4-antidipyrine and remantadine was studied on a biological model of influenza infection (intranasal infection of non-linear mice with influenza A virus (Aichi / 2/28)), as well as a combination proper use of these drugs.

Example 1 of a specific experiment. The effect of the individual components of the pharmaceutical composition of iodantipyrine and remantadine on the course of the flu in the experiment was investigated. For this, the animals were divided into groups receiving iodantipyrine at doses of 12.5, 25, 50, 100, 250 and 500 mg / kg and remantadine at doses of 25, 12.5, 6.25 and 3.0 mg / kg. The control group consisted of untreated animals infected with the influenza virus. The indicators of the effectiveness of the treatment were: a decrease in the mortality rate of mice compared to the control group during the period of maximum death, as well as an increase in the average duration of the incubation period. During the experiment, it was shown that in the control group, the survival rate was 0%, and the average incubation period was 6.6 days. In mice treated with 4-iodantipyrine, the maximum protective effect (survival rate of 33.3% of animals) was observed with therapeutic and prophylactic administration at a dose of 250 mg / kg (Table 1). A higher concentration (500 mg / kg) of 4-iodantipyrine did not enhance the protective effect. Concentrations of 4-iodantipyrine of 100 mg / kg or less did not provide reliable protection of animals from influenza infection compared with the control group. sing. Remantadine at a dose of 25 mg / kg showed survival only

17.4%, and even lower doses were completely ineffective, while the average incubation period decreased in parallel (Table 1). Example 2 of a specific experiment.

The therapeutic and prophylactic effect of the combined use of doses of 4-iodo-antipyrine and remantadine was also investigated in a model of experimental influenza infection. Animals of the experimental groups received combined treatment with iodantipyrine and remantadine in the following doses: 100 and 25 mg / kg; 50 and 12.5 mg / kg; 25 and 6.25 mg / kg; 12.5 and 3 mg / kg; 6 and 1.5 mg / kg; 3 and 0.75 mg / kg, respectively. The best effect was observed with the combined administration of 4-iodo-antipyrine and remantadine at doses of 100 and 25 mg / kg, respectively, which ensured the survival of 75% of the animals, exceeding by 4 times the percentage of protection for each drug in the indicated doses separately. Mutual two-fold reduction in the doses of the components of the combination gave the same protective effect. The minimum effective concentrations of 4-iodantipyrine and remantadine, which together provide 50% protection, were 12.5 and 3.0 mg / kg, respectively. Consequently, with combined administration, the dose of drugs decreased by 10–16 times compared with separate use, and the effectiveness was disproportionately higher.

Example 3 of a specific experiment.

To compare the effectiveness of various administration schemes, the therapeutic and prophylactic effect of the combination of iodantipyrine and remantadine in the following doses was studied separately: 100 and 25 mg / kg; 50 and 12.5 mg / kg; 25 and 6.25 mg / kg; 12.5 and 3 mg / kg; 6 and 1.5 mg / kg; 3 and 0.75 mg / kg, respectively (table. 3). When administered 48, 24, 1 hour before infection (prophylactic regimen) or 24, 48, 72 hours after infection (treatment regimen), 4-dodipyrine and remantadine also provided solid protective effect. The maximum effect - the survival rate of 83.3%, the average duration of the incubation period of 9.3 days with the treatment regimen and 91.7% survival, 9.8 days, the duration of the incubation period of the prophylactic regimen was observed with the same dose ratio of iodine pyrine and remantadine: 100 and 25 mg / kg, respectively. Comparing percentages and the average duration of the incubation period when using similar doses of 4-dodipyrine (100 mg / kg) and remantadine (25 mg / kg), a prophylactic regimen should be preferred.

We also studied the effect of the treatment as separate components of iodantipyrine and remantadine, and their combined use in doses of 100 and 25 mg / kg on some immunological parameters: the concentration of influenza virus (Ig EID ₅ o / 0.2 ml), serum interferon ( IU / ml) and the level of antihemagglutinins (log 2).

Example 4 of a specific experiment.

The presence of influenza virus in the lungs of mice was determined on the 1st, 3rd, 5th, 7th day from the moment of infection. For this purpose, lungs were taken from 5 mice and 10-11-day-old chicken embryos were infected with the prepared 20% suspension. Virus concentration was evaluated in Ig EIDso / 0.2 ml. At the same time, the blood serum of experimental mice was examined for the presence of interferon and antihemagglutinins. Interferon activity (U / ml) was determined by titration of blood sera in a Z-929 cell culture versus 100 TCD ₅ o / O, l ml of air force in accordance with generally accepted methods and the content of antihemagglutinins.

It was shown that the maximum accumulation of influenza virus in the lungs of mice was observed on the 3rd day after infection (table. 4). By the 5th day, the virus content decreased to 3.5 units, and on the 7th day during the peak of influenza pneumonia and maximum death of animals, the titer the virus amounted to 1.5 units The obtained results confirm the data of other researchers on the maximum concentration of influenza virus in the lungs in the early stages of infection, followed by a decrease due to the development of an active inflammatory process in the lungs. Separate therapeutic administration of 4-dodipyrene (100 mg / kg) and remantadine (25 mg / kg) did not significantly affect the dynamics of the influenza virus in the lungs of mice (titer reduction by 0.5-1.0 units). The combined administration of drugs in the same doses led to a decrease in the virus content in the lungs by 2.5 units. by 3-5 days of the experiment and by the termination of virus isolation on the 1st and on the 7th day.

Serum interferon was detected in mice of the control group (infected with the virus), starting from the 1st day of the experiment, with a maximum content (80 U / ml) by the 3rd day, and from the 5th day of infection its content sharply decreased. A similar pattern was observed with the separate administration of 4-iodo-antipyrine (100 mg / kg) and remantadine (25 mg / kg). The introduction of the pharmaceutical composition (4-iodantipyrine at a dose of 100 mg / kg, remantadine at a dose of 25 mg / kg) contributed to a more stable production of interferon during the 7 days of the experiment, which can be a fairly good prognostic sign (Table 5). The combined administration of 4-dantipyrine at a dose of 100 mg / kg with remantadine at a dose of 25 mg / kg (according to the treatment regimen) was accompanied by a higher level of antihemagglutinins (6.0 log 2) by 7 days of infection: slightly lower (5.0 log 2) there was this indicator in the treatment with only 4-iodantipyrine (100 mg / kg), and the antibody content in the treatment with rimantadine only (25 mg / kg) did not differ from that in the control group of infected animals that were not subjected to treatment (Table 6).

Industrial applicability The inventive antiviral agent can be widely used as a pharmaceutical composition for the treatment and prevention of influenza.

An antiviral agent has a pronounced antiviral activity in therapeutic and prophylactic regimens. When compared with the individual components, the proposed tool is a pharmaceutical composition containing iodantipyrine and remantadine in the following ratios, May. %: iodantipyrine - 23 - 97, remantadine - 3 - 67, surpasses them in efficiency. In addition, reducing the dose of remantadine can reduce the occurrence of side symptoms.

Table 1

The effect of various doses of 4-iodo-antipyrine and remantadine on the course of experimental influenza infection in non-inbred mice with a therapeutic and prophylactic regimen

Note: Here and in table. 2, 3 * - the differences are significant compared with the control at Pt and Pu <0.05

table 2

The effectiveness of the combined action of 4-iodantipyrine and remantadine in the treatment-and-prophylactic regimen of administration to mice in experimental influenza infection

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Table 3

The efficacy of the combined action of 4-iodantipyrine and remantadine in the treatment and prophylactic regimen of administration to mice against the background of an experimental influenza infection

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Table 4

The content of influenza virus in the lung tissue of mice treated separately and combined with 4-dodipyrine (100 mg / kg) and remantadine (25 mg / kg)

Table 5

The content of interferon in the blood serum of mice treated separately and combined with 4-dodipyrine (100 mg / kg) and remantadine (25 mg / kg)

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Table 6

The content of anti-hemagglutinins in the blood serum of mice treated separately and combined with 4-i-antipyrine (100 mg / kg) and remantadine (25 mg / kg)

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Sources of information from the prior art 1. Mashkovsky M. D. Medicines. - M .: Medicine, 1986, v. 2.

5 2. RF patent Ns 2025126 of 02.25.2003 "Means for the treatment and prevention of tick-borne encephalitis induced in the experiment." 3. RF patent JUfe 2175548 of 04/20/1998 "Means for the prevention of influenza and other acute respiratory viral infections", 4. 4. RF patent JN ° 2145218 of 02/10/2000 "Pharmaceutical composition having antiviral effect."

5. RF patent N ° 2229877 dated 06/10/2004 "Means for the treatment of influenza."

6. RF patent N ° 2071323 dated 01/10/1997 "Antiviral drug 15" Polyrem ""

7. RF patent JYe 2185822 from 07.27.2002. "The drug for the treatment and prevention of viral diseases."

8. Ilyenko V. I. Methods of testing and evaluating the antiviral activity of chemical compounds against influenza virus: 0 Guidelines. - L., 1977.

Claims

CLAIM

An antiviral agent containing rimantadine, characterized in that it additionally contains iodantipyrine in the following ratio of components, wt.%: Remantadine - 3 - 67 Iodantipirin - 23 - 97