RU2394618C2 - Water-soluble polymeric complexes of arbidol - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to high-molecular compounds for medical purposes. Water-soluble polymeric complexes of antiviral agent arbidol of general formula:

are described, where: Arb - arbidol: ethyl ester 6-bromo-4-dimethyl-aminometil-1-methyl-5-oxy-2-feniltiometilindolinil-3-carboxylic acid hydrochloride monohydrate; m1=100-(m2+m3) mol %; m2=(7.6-9.8) mol %; m3=(11.5-13.6) mol %; content of Arb=26.4-32.1 wt %.

EFFECT: obtained water-soluble polymeric complexes of arbidol may find application in pharmacology, as they can serve as basis for new effective and safe antiviral drugs and their dosage forms.

1 cl, 6 ex, 4 tbl, 2 dwg

Description

The invention relates to macromolecular medical compounds, namely to water-soluble polymer complexes containing the drug arbidol, of the General formula

where Arb is arbidol: ethyl 6-bromo-4-dimethyl-aminomethyl-1-methyl-5-hydroxy-2-phenylthiomethylindolinyl-3-carboxylic acid hydrochloride monohydrate;

m ₁ = 100- (m ₂ + m ₃ ), mol.%;

m ₂ = (7.6-9.8) mol%;

m3 = (11.5-13.6) mol%

with M.M. (19.000-31.000) Da.

The Arb content is 26.4-32.1 wt.%.

As a result of complexation of arbidol with polymers having a molecular weight of 19,000-31,000 Da and consisting of copolymers of acrylamide (AA) and 2-acrylamido-2-methylpropanesulfonic acid (AAMPSK), a number of water-soluble derivatives of arbidol have been obtained, which have reduced toxicity compared to arbidol, increased bioavailability , improved pharmacodynamics, and these polymer complexes fully maintain a high level of biological activity of arbidol, in particular antiviral activity. The content of arbidol in the resulting water-soluble polymer complexes is 26.4-32.1 wt.%.

The created water-soluble polymer complexes of arbidol can be used in pharmacology, as they can serve as the basis for new effective and safe antiviral drugs and their dosage forms.

The need to have in the arsenal of drugs highly effective antiviral drugs with a wide spectrum of activity against influenza A and B viruses and other acute respiratory viral infections (ARVI) leads to a modification of the formulas of the drugs found.

1. The reason for the modifications is the high toxicity of many drugs used or their insufficient solubility.

Commonly used etiotropic antiviral drugs are remantadine, neuraminidase inhibitors - zanamivir and oseltamivir, ribavirin. The use of remantadine is limited by its lack of activity against influenza B viruses, as well as by the widespread use of resistant strains of the influenza virus in recent years [CDC Health Alert. CCID .: www.cdc.gov/flu/han011406]. Regarding neuraminidase inhibitors, it can be noted that along with the high cost of these drugs, their disadvantages include side effects in the form of irritation of the nasopharynx when taking zanamivir and in the form of nausea and vomiting when taking oseltamivir. In addition, they are effective only at the very beginning of a viral infection. Often, in the treatment of severe forms of acute respiratory viral infections, ribavirin and its domestic analogue, ribamidil, are also used. Due to the high toxicity, these drugs are used only in a hospital setting for people with weakened immune systems [Kiselev O.I., Deeva E.G., Platonov V.G., Ilyenko V.I. Antiviral drugs // "Flu and other SARS." - SPb. - 2003. - S.96-146].

2. One of the effective and technological ways to reduce the toxicity of drugs while maintaining specific biological activity is their complexation with water-soluble non-toxic ionic carrier polymers [Eropkin M.Yu., Solovsky MV, Eropkina EM, Shultseva E.L. . Comparative study of the cytotoxic effect of polymer derivatives of aminoglycoside antibiotics // Toxicol. messenger. - 2006. - No. 5. - S.18-22]. The polymer preparations of remantadine (its complex with sodium alginate - algir and the complex with a copolymer of N-vinyl succinimide with vinyl amidosuccinic acid - polymer) have a wider spectrum of antiviral activity compared to remantadine. An important factor is the prolonged effect of polymer derivatives of remantadine, which promotes prolonged circulation of the antiviral component in effective therapeutic doses [Kiselev OI, Deeva EG, Slita AV, Platonov VG Antiviral drugs for the treatment of influenza and acute respiratory infections. Design of drugs based on polymer carriers. "Time". St. Petersburg, 2000].

3. Arbidol - ethyl 6-bromo-4-dimethylaminomethyl-1-methyl-5-hydroxy-2-phenylthiomethylindolinyl-3-carboxylic acid hydrochloride monohydrate, which has a broad spectrum of action against viruses, is known and widely used as an antiviral agent influenza A and B and other acute respiratory viral infections (ARVI)

Arbidol, in addition to its antiviral effect, is an immunomodulator, an inducer of interferon and has antioxidant properties [Guskova T.A., Glushkov R.G. Arbidol - immunomodulator, interferon inducer, antioxidant. TsHLS-VNIHFI, M., 1999, 2001]. The virus-inhibiting effect of arbidol is determined by its ability to inhibit the fusion of the lipid membrane of the virus with endosome membranes, which occurs inside the cells. It was shown that arbidol does not affect the adsorption, transcription and translation, as well as the neuraminidase activity of the influenza virus [I. Leneva The mechanism of the virus-specific action of the drug arbidol. Abstract of the doctor. Diss., St. Petersburg, 2005]. Currently, arbidol is widely distributed in the domestic pharmaceutical market and is one of the main etiotropic anti-influenza drugs.

At the same time, a significant drawback of the drug is that it is practically insoluble in water, which reduces its bioavailability and limits the possibility of creating new dosage forms based on it (solutions, aerosols, hydrophilic ointments). In addition, arbidol as a therapeutic drug is effective only in the early stages of the flu disease - no more than 2 days from the onset of the disease [Leneva IA, Guskova TA, Glushkov R.G. Medicines for chemotherapy and chemoprophylaxis of influenza: features of the mechanism of action, effectiveness and safety // Chem. - farm. Journal. - 2004. - No. 11. - S.8-14].

With intragastric administration, arbidol refers to low-toxic drugs [Guskova TA., Glushkov R.G. Arbidol - immunomodulator, interferon inducer, antioxidant. TsHLS-VNIHFI, M., 1999, 2001], however, when administered parenterally in mice and rats, its DL ₅₀ is 109 and 140 mg / kg, respectively, which allows it to be classified as moderately toxic drugs [Berezovskaya I.V. Classification of chemicals according to the parameters of acute toxicity with parenteral routes of administration // Chem.-pharm. g. - 2003. - T. 37, No. 3. - S. 32-34]. When evaluated in cell cultures, its IC ₅₀ (average inhibitory concentration, which reduces cell viability by 50%) is 40-60 μg / ml [I. Leneva The mechanism of the virus-specific action of the drug arbidol. Diss. Doct. biol. Sciences, St. Petersburg, 2005]. In addition, according to our data, when the drug is stored for about 6 months, its cytotoxicity increases significantly (IC ₅₀ decreases to 20 μg / ml on an MDCK cell culture). Moreover, the pharmacological index, or selectivity index (the ratio between the cytotoxic concentration and the average virus-inhibiting concentration) is only 3.75, which is an unacceptable indicator for such a widely used drug (as a rule, the selectivity index should at least be greater than 10) [Khabriev R .U. (ed.). Guidelines for the experimental (preclinical) study of new pharmacological substances. - M., 2005].

4. The technical task and the positive result of the invention is the creation of water-soluble derivatives of arbidol with reduced toxicity while maintaining a wide range and a high level of biological activity, in particular antiviral activity.

This task is achieved due to the complexation of arbidol with copolymers of acrylamide (AA) with 2-acrylamido-2-methylpropanesulfonic acid (AAMPSK), resulting in a polymer complex

where Arb is arbidol: ethyl 6-bromo-4-dimethyl-aminomethyl-1-methyl-5-hydroxy-2-phenylthiomethylindolinyl-3-carboxylic acid hydrochloride monohydrate;

m ₁ = 100- (m ₂ + m ₃ ), mol.%;

m ₂ = (7.6-9.8) mol%;

m3 = (11.5-13.6) mol%

with M.M. (19.000-31.000) Da, and the content of arbidol in it (26.4-32.1) wt.%.

5. The invention is the use of copolymers of AA with AAMPSK for complexation of arbidol, as a result of which, with strictly selected proportions of arbidol and copolymers, water-soluble polymer complexes with an arbidol content of 26.4 to 32.1 wt.%, The hydrochloride and monohydrate in the composition of arbidol are included in the structure of the link m ₂ . The content of units in the polymer chain in molar% is: m ₁ (AA) - [100- (m ₂ + m ₃ )], m ₂ (Arb complex with AAMPSK) - (11.5-13.6).

Small in magnitude (19.000-31.000 Da) molar masses of polymers (MM) ensure their complete elimination from the body by the mechanism of renal filtration, which, in the absence of biological activity in sulfonated copolymers, leads to the formation of a complex in a water-soluble form acceptable to the body while maintaining the biological activity of arbidol as such.

Polymeric derivatives of arbidol and, in particular, the claimed water-soluble polymer complexes are not described in the literature.

The choice of AA-AAMPSC copolymers as modifiers of the properties of arbidol is due to the following reasons:

a) These copolymers, like all polymers of acrylamide (AA), have highly hydrophilic polymer chains, are highly soluble in water.

b) The copolymers contain strongly acid sulfo groups and in aqueous solutions easily form a salt bond with primary, secondary and tertiary amines, namely arbidol.

c) According to the literature [M.V. Solovskiy, M.Yu. Eropkin, U.M. Eropkin and others. Synthesis and properties of low molecular weight copolymers of acrylamide with 2-acrylamido-2-methylsulfonic acid - potential carriers of biologically active substances // J. applied chemistry. - 2008, - T. 80. - issue 10. - C.1674-1678] copolymers AA-AAMPSK (≤ 23 mol.% Sulfo groups) are non-toxic in vitro.

6. Preparation of carrier polymers and complex formation of arbidol with them

Carrier polymers - copolymers of acrylamide with 2-acrylamido-2-methylpropanesulfonic acid (AA-AAMPSK) were obtained by heterophase radical copolymerization of co-monomers in isopropanol [M.V. Solovsky, M.Yu. Eropkin, E.M. Eropkina, etc. J. applied chemistry. - 2007. - T. 80. - Issue 10. - S.1674-1678]. The composition of AA-AAMPSK copolymers was calculated according to the elemental analysis for sulfur content. The molecular weights of AA-AAMPSC copolymers were determined by the viscometric method using the Mark-Kun-Hauwink equation, known for polyacrylamide [Encyclopedia of Polymers. Publishing House "Soviet Encyclopedia". 1972. T. 1. - P.30].

The structure of the copolymers was confirmed by titration of sulfo groups, as well as by IR spectra, in which, in contrast to the IR spectrum of poly-AA, new characteristic absorption bands of -SO ₃ H groups are observed in the region of 1227 cm ^-1 , 1040 cm ^-1 , 624 cm ^{- 1} .

The complexation of copolymers AA-AAMPSK with arbidol was carried out in water at room temperature, with a mass ratio of copolymer / arbidol equal to 2.3-2.5 / 1. AA-AAMPSC copolymers were dissolved in distilled water at a rate of 3.0-4.0 mg / ml and dry arbidol (26-30 mg) was added to the solution with stirring. The mixture was stirred for 40-60 minutes, while all arbidol completely passed into solution. The reaction mixture was filtered. Polymer complexes were isolated by freeze drying. The arbidol content in the obtained polymer complexes was determined by UV spectroscopy. An intense absorption band of arbidol was used with a maximum at 320 nm (ε = 13.700 mol ^-1 · cm ^-1 ). UV spectra were recorded in a mixture of DMF + H ₂ O (volume ratio 1: 9). The complexation of the copolymers with arbidol is shown schematically in the diagram:

Tests of the resulting complexes were carried out in vitro.

7. Determination of antiviral activity of drugs

The antiviral effect of the synthesized compounds and preparations was determined against the reference strain of human influenza virus A / Victoria / 35/72 (H3N2). The action of arbidol and its polymer complex was also evaluated in relation to the highly pathogenic strain of bird flu isolated and deposited at the research institute of influenza - A / duck / Kurgan / 8/05 (H5N1).

The presence of the virus in the incubation medium was determined by the micromethod of the hemagglutination reaction with a 0.5% suspension of human erythrocytes of I (0) blood group. Virus titer was expressed in decimal logarithms of 100 μl (logTID ₅₀ ). Antiviral activity of the samples was evaluated by reducing the titer of the virus in the experimental wells of the tablets compared with the control (ΔlgTID ₅₀ ). The average virus-inhibiting concentration of the sample (VIC ₅₀ ) was calculated by the cytopathic response of the cells when exposed to a specific titer virus, which was evaluated in the microtetrazolium test. Microtetrazolium test (MTT) was performed according to Mosmann [Mosmann T. J Immunol Meth, 1983, v. 65 (1), p. 55-63]. Cell cultures grown in 96-well plates were incubated for 3 hours in MTT solution (0.5 mg / ml) in buffered saline at 37 ° C in a CO ₂ incubator. The resulting water-insoluble crystals of formazan were extracted with 95 ° ethanol for 30 min and the optical density of the samples was measured at a wavelength of 550 nm on a Chameleon plate analyzer (Hydex, Finland).

The degree of inhibition of cell viability in culture correlates with the development of viral infection in vitro.

Antiviral activity was also evaluated against herpes simplex virus type I HSV1 / 248/88 and adenovirus type III Ad / 3 / et / 4120 in cell culture A-549 (human lung carcinoma line). The drugs were added according to the treatment-and-prophylactic regimen — 30 minutes before the virus suspension was introduced into the culture. The cytopathic reaction was taken into account after 48 and 72 hours according to the degree of degradation of the monolayer (microscopic examination of the culture), as well as by the MTT method. In the latter case, the regression method was used to calculate the VIC ₅₀ .

8. Determination of cytotoxicity of the studied compounds in vitro

The toxicity of arbidol (Masterlek, M., capsule form), carrier polymers and polymer complexes of arbidol was determined by incubation of MDCK cell cultures with successive dilutions of the preparations in Igla-MEM serum-free medium. The standard incubation time with the preparations was 72 hours. In vitro methods of assessing toxicity were cell restoration in a culture of the fluorescent dye resazurin (Sigma, USA) [Clothier R., Starzec G., Pradel L. et al. The prediction of human skin responses by using the combined in vitro fluorescein leakage / Alamar blue (resazurin) assay // ATLA. - 2002. - V. 30. - P.493-504] or MTT tetrazolium dye - 3- (4,5-dimethylthiazolyl-2) 2,5-diphenyltetrazolium bromide (ICN Pharmaceuticals, USA). The MTT method was carried out as described above.

Both methods reflect the integral activity of mitochondrial dehydrogenases and serve as adequate indicators of cell viability in culture and the intensity of oxidative processes. The dye resazurin (in the English language literature - Alamar Blue), when reduced by mitochondrial dehydrogenases, turns into the fluorescent product resarufin (λ _max excitation = 530 nm, λ _max emission = 590 nm). The technique is adapted for 96-well plates: the measurements were carried out in a Chameleon plate analyzer with appropriate interference filters for fluorescence. The advantage of the method is that fluorescence is measured directly in the wells after incubation with dye without additional extraction, since the reaction product, resarufin, is freely excreted from the cells into the medium. The cells then remain viable, and on the same plate after washing the dye, another test can be carried out, for example, binding of neutral red, determination of DNA or protein, etc. [Clothier R., Starzec G., Pradel L. et al. The prediction of human skin responses by using the combined in vitro fluorescein leakage / Alamar blue (resazurin) assay // ATLA. - 2002. - V. 30. - P.493-504].

An IC ₅₀ , a cytotoxic dose causing a 50% drop in the corresponding parameter (intact cells), which was calculated using dose-response linear regression equations, was taken as a criterion of toxicity in both methods. At least 4 points with three or more independent repetitions of the experiment were taken for each concentration. We used two options for representing concentration values - linear and logarithmic. Mathematical and statistical data processing was performed in Excel 2000.

The invention is presented in the examples.

Example 1. 68 mg of a copolymer AA-AAMPSK containing 19.1 mol.% Sulfo groups, with MM 19.000 Da, was dissolved in 20 ml of distilled water. To the resulting solution, 27.2 mg of dry arbidol was added with stirring. The mass ratio of copolymer / arbidol is 2.5: 1. The mixture was vigorously stirred at room temperature for 40 minutes. In this case, arbidol completely passed into solution. The target solution was filtered through a solid Shot filter (40 pores). The filtrate was frozen and subjected to freeze drying. Received 75.1 mg (78.8%) of a water-soluble polymer containing 28.1 wt.% Arbidol. The composition and biological characteristics of the obtained product are given in Tables 1-4 and in Figs. 1 and 2. It can be seen from Table 1 that the antiviral activity of the arbidol complex with this carrier is comparable with the antiviral activity of unmodified arbidol in relation to the model strain of influenza A virus (H3N2 ), and its in vitro toxicity is about 10 times lower than that of unmodified arbidol (the corresponding IC ₅₀ is an order of magnitude higher). According to the data presented in Table 2, it follows that arbidol has a distinct antiviral dose-dependent activity (ΔlogTID ₅₀ > 2.0) in this system both in relation to model virus A (H3N2) and highly pathogenic bird flu A (H5N1 ), and the activity of the complex with respect to the latter is significantly higher than that of unmodified arbidol. A / duck / Kurgan / 8/06 virus (H5N1) was isolated on MDCK cell culture and transferred to chicken embryos (C2 / E1). Hemagglutinating activity of the virus: 32-128 GAE / 0.2 ml, infectious activity: 6.0 TID ₅₀ / 0.2 ml.

More detailed data on the effects of polymer-modified arbidol are presented in figures 1 and 2. Figure 1 shows the antiviral activity of arbidol (dashed line) and the Arb-AA-AAMPSK complex (solid line) against highly pathogenic avian influenza A / duck / Kurgan / 8/05. On the abscissa axis is the concentration of arbidol and complex in μg / ml. The ordinate axis - wholesale. pl. at 550 nm (MTT). Figure 2 shows the cytotoxicity of arbidol (dashed line) and the Arb-AA-AAMPSK complex (solid line) in MDCK cell culture. Designations are the same as in figure 1.

ΔlogTID ₅₀ = 2.0 for arbidol at a concentration of 6.25 μg / ml. At 10 TID _{50, the} virus-inhibiting concentration (VIC ₅₀ ) of arbidol was VIC ₅₀ = 5.2 μg / ml. In vitro average toxic concentration of arbidol in relation to MDCK cells: IC ₅₀ = 19.5 μg / ml (see page 2). Thus, the pharmacological index (drug selectivity index) at 10 TID ₅₀ is 3.75, which is clearly an unsatisfactory indicator for such a widely used drug.

At the same time, for polymer-modified arbidol in equimolar concentration (taking into account the mass content of arbidol in it) at 10 TID ₅₀ VIC ₅₀ = 12.5 μg / ml, and the toxicity of IC ₅₀ = 190 μg / ml. Thus, the selectivity index for this drug was 15.2 under the same conditions, which is four times better than that of unmodified arbidol.

From this we can conclude that polymer-modified arbidol not only exhibits high antiviral activity against the highly pathogenic strain of influenza A virus (H5N1) isolated in Russia, but also has significantly lower toxicity and a much higher pharmacological index compared to unmodified arbidol.

Table 3 shows the antiviral effect of arbidol and its complex with AA-AAMPSK (example No. 1 from table 1) in relation to the herpes simplex virus HSV1 / 248/88 in cell culture A-549. A significant antiviral effect of arbidol and a comparable dose-dependent effect of its complex with AA-AAMPSK against herpes simplex virus type 1 have been established. The antiviral effect is comparable to that of the human influenza virus.

Table 4 shows the antiviral effect of arbidol and its complex with AA-AAMPSK (example No. 1 from table 1) in relation to adenovirus A <3/3 / et / 4/20 in cell culture A-549. A weak antiviral effect of both arbidol and its polymer-modified derivative on adenoviruses was noted. In this case, the concentration dependence of the action of the latter is not observed. The effect of arbidol when it is evaluated by two methods is the same, while for the polymer derivative it is lower when evaluated by MTT reduction.

Example 2. In the conditions of example 1 of 68 mg of a copolymer AA-AAMPSK containing 22.0 mol.% Sulfo groups, with M.M. 31,000 Da and 27.2 mg of arbidol (weight ratio copolymer / arbidol 2.5: 1) obtained 79.2 mg (83.2%) of a water-soluble polymer containing 28.6 wt.% Arbidol. The composition and biological characteristics of the obtained product are given in table 1.

Example 3. In the conditions of example 1 of 68 mg of a copolymer AA-AAMPSK containing 22.8 mol.% Sulfo groups, with M.M. 28,000 Da and 27 mg of arbidool (mass ratio copolymer / arbidol 2.5: 1) obtained 79.4 mg (83.6%) of a water-soluble polymer containing 26.4 wt.% Arbidol. The composition and biological characteristics of the obtained product are given in table 1.

Example 4. In the conditions of example 1 of 68 mg of a copolymer AA-AAMPSK containing 22.8 mol.% Sulfo groups, with M.M. 28,000 Da and 30.4 mg of arbidol (weight ratio copolymer / arbidol 2.3: 1) obtained 88.3 mg (87.9%) of a polymer containing 32.1 wt.% Arbidol. The composition and biological characteristics of the obtained product are presented in table 1. As can be seen from the table, the antiviral activity of all polymers obtained in examples No. 1-4 is approximately the same. The corresponding indicator (ΔlogTID ₅₀ ) exceeds 2.0, which indicates a rather high antiviral activity of the synthesized polymer complexes. The antiviral activity of all the complexes obtained in Examples Nos. 1-4 is not statistically different both from each other and from the activity of unmodified arbidol (non-parametric Mann-Whitney test). The in vitro toxicity of all the complexes studied in Examples 1-4 is also approximately the same. A slight variation in cytotoxicity (statistically unreliable) is observed depending on the content of sulfo groups. At the same time, the toxicity of all synthesized polymer complexes is almost an order of magnitude lower than the in vitro toxicity of unmodified arbidol, which we determined under the same experimental conditions (Table 1).

Example 5 (control). In the conditions of example 1 of 68 mg of a copolymer AA-AAMPSK containing 22.8 mol.% Sulfo groups, with M.M. 28,000 Da and 22.7 mg of arbidol (weight ratio copolymer / arbidol 3.0: 1), 76.5 mg (84.4%) of a water-soluble polymer containing 24.8 wt.% Arbidol was obtained. The composition and biological characteristics of the obtained product are given in table 1.

This complex had a low antiviral activity, which corresponds to the lowest mass content of arbidol in it compared to other complexes (examples 1-4).

Example 6 (control). 70 mg of a copolymer AA-AAMPSK containing 19.1 mol.% Sulfo groups with M.M. 19,000 Da, dissolved in 20 ml of water. To the resulting solution was added 36 mg of arbidol (weight ratio copolymer / arbidol 1.95: 1). In this case, arbidol did not dissolve in the copolymer solution.

Analysis of tabular data allows us to draw the following conclusions:

1. The inventive task has been solved by the claimed water-soluble polymer complexes of arbidol with low molecular weight copolymers of acrylamide with 2-acrylamido-2-methylpropanesulfonic acid.

2. The antiviral activity of the claimed polymer complexes in relation to all viruses we studied (human influenza virus A (H3N2), highly pathogenic avian influenza virus A (H5N1), herpes simplex virus type I (HSV1), type III adenovirus) is comparable with the antiviral activity of unmodified arbidol.

3. In vitro toxicity of the claimed complexes is approximately an order of magnitude lower than toxicity under the same conditions of unmodified arbidol.

4. The pharmacological index (selectivity index) calculated for the polymer complex (Example No. 1) with respect to influenza A virus (H5N1) was 15.2 versus 3.75 for unmodified arbidol. Thus, the pharmacological index of the polymer derivative is four times higher than the corresponding indicator of unmodified arbidol.

5. The declared interval parameters are determined by the following factors:

a) The molecular weight range of the AA-AAMPSK 19.000-31.000 Da copolymers, firstly, provides a solution to the inventive task, and secondly, ensures the complete removal of the carrier polymer from the body by the renal filtration mechanism.

b) The claimed content of arbidol in the complex (26.4-32.1) wt.% ensures the solubility of the claimed complexes and their high antiviral activity. When the arbidol content in the complex is 24.8% (Example 5), the complex is characterized by lower antiviral activity. A water-soluble complex with a desired arbidol content of 34% (Example 6) cannot be obtained.

Table 3 The antiviral effect of arbidol and its complex with copolymer AA-AAMPSK (example No. 1 from table 1) in relation to the herpes simplex virus HSV1 / 248/88 in the cell culture A-549. The drugs were introduced 30 minutes before infection with the virus. Analysis of results after 72 hours. Evaluation of the cytopathogenic effect (CPD) by morphological changes in cells (light-optical microscopy) and restoration of MTT dye. Average 3-4 independent experiments Indicator The control Arbidol, 10 mcg / ml Ar-AA-AAMPSK, 33 * μg / ml Ar-AA-AAMPSK, 67 μg / ml CPD, (logTID ₅₀ ), microscopic. morphology assessment 3.67 1.83 2.5 2.13 Antiviral effect on micr. evaluation (ΔlogTID ₅₀ ) 1.84 1.17 1,54 CPP (logTID _5o), Restore. MTT 2.6 0.55 0.95 0.4 The antiviral effect of the recovery. MTT (ΔlоgТИД ₅₀ ) 2.1 1.65 2.2 * - see the signature to table.2.

Table 4 The antiviral effect of arbidol and its complex with copolymer AA-AAMPSK (example No. 1 from table 1) in relation to adenovirus Ad / 3 / et / 4/20 in cell culture A-549. The remaining conditions are as in table 3 Indicator The control Arbidol, 10 mcg / ml Arb-AA-AAMPSK, 33 μg / ml Arb-AA-AAMPSK, 67 mcg / ml CPD, (logTID ₅₀ ), microscopic. morphology assessment 3.0 2.25 2.13 2.25 Antiviral effect on micr. assessment (ΔlоgТИД ₅₀ ) 0.75 0.87 0.75 CPD, (logTID ₅₀ ), recovery MTT 2.04 1.3 1,5 1.9 The antiviral effect of the recovery. MTT (ΔlogTID ₅₀ ) 0.74 0.54 0.14

Claims (1)

Water-soluble polymer complexes of the antiviral arbidol of the general formula:

where Arb is arbidol: ethyl 6-bromo-4-dimethyl-aminomethyl-1-methyl-5-hydroxy-2-phenylthiomethylindolinyl-3-carboxylic acid hydrochloride monohydrate;
m1 = 100- (m2 + m3) mol%;
m2 = (7.6-9.8) mol%;
m3 = (11.5-13.6) mol%;
Arb content = 26.4-32.1 wt.%.

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