RU2806534C1 - Adaptogenic, immunomodulating and anti-viral pharmaceutical composition of oxyethylammonium methylphenoxyacetate - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to a solid pharmaceutical composition having adaptogenic, immunomodulatory and antiviral properties against influenza A, influenza B, parainfluenza viruses and seasonal coronavirus OC43. The following is proposed: solid pharmaceutical composition with adaptogenic, immunomodulatory and antiviral properties against influenza A, influenza B, parainfluenza and seasonal coronavirus OS43 viruses, containing oxyethylammonium methylphenoxyacetate, additional components and excipients in the following ratio of components, wt.%: 20–60 of oxyethylammonium methylphenoxyacetate, 0.00001–0.5 of o-cresol and/or 0.00001-0.5 of 2-phenoxyacetic acid and/or 2-(3-methylphenoxy) acetic acid and/or 2-(4-methylphenoxy) acetic acid or a mixture of these substances, 0.00001–0.29 of phosphorus and/or 0.0000001–0.00050 of zinc or a mixture thereof, up to 100 of microcrystalline cellulose, lactose monohydrate, sodium stearyl fumarate, calcium stearate, aerosil, potato starch or mixtures thereof.

EFFECT: above invention has increased adaptogenic, immunomodulatory and antiviral effects, increased bioavailability and reduces negative side reactions.

4 cl, 4 dwg, 15 tbl, 3 ex

Description

Field of technology to which the invention relates

The invention relates to the field of medicine, namely to the pharmaceutical industry, and can be used for the production of a drug in solid dosage form based on oxyethylammonium methylphenoxyacetate with additional agents, which has adaptogenic, immunomodulatory and antiviral effects.

State of the art

Immunostimulant with a wide spectrum of biological activity - INN oxyethylammonium methylphenoxyacetate (tris(2-hydroxyethyl)ammonium 2-methylphenoxyacetate; tris(2-oxyethyl)ammonium orthocresoxyacetate) formula:

- was synthesized in the early 1970s at the Irkutsk Institute of Chemistry. A.E. Favorsky SB RAS (formerly the Irkutsk Institute of Organic Chemistry of the USSR Academy of Sciences) to domestic scientist M.G. Voronkov (author's certificate SU No. 515742, filing date: 08/06/1973).

Hydroxyethylammonium methylphenoxyacetate was studied for a long time and only in August 1994 the Russian Ministry of Health was approved for medical use under the name Trecrezan® as an adaptogenic agent with a broad spectrum of action, and was also protected by the patent RU2063749 “A MEDICINE WITH AN ADAPTOGENIC ACTION” (priority date: 02.24. 1994).

Currently, the drug Trekrezan® (INN hydroxyethylammonium methylphenoxyacetate, LSR-008909/09 Grotex LLC) belongs to the group of adaptogenic drugs. The drug stimulates the production of α- and γ-interferons, which determines the spectrum of its biological activity (immunostimulating, adaptogenic), affects the immune status of the body due to the activation of cellular and humoral immunity, and stimulates the phagocytic activity of macrophages. The drug strengthens the body's immune system, increases endurance during physical and mental stress, reduces the effect of various toxins, increases the body's resistance to hypoxia, low and high temperatures and other adverse environmental factors.

The drug Trekrezan® is now available on the Russian market, having the following composition for 1 tablet (composition 1) :

hydroxyethylammonium methylphenoxyacetate 0.2 g;

Excipients:

potato starch 0.0925 g;

lactose monohydrate (milk sugar) 0.2 g;

sodium stearyl fumarate 0.005 g;

colloidal silicon dioxide (aerosil) 0.0025 g.

In addition, patent RU 2762552 (priority date: 10/05/2020) was recently issued for a pharmaceutical composition of hydroxyethylammonium methylphenocyacetate with the following composition (composition 2) :

hydroxyethylammonium methylphenoxyacetate 0.2 g;

Excipients:

mycrystalline cellulose 0.0925 g;

lactose monohydrate (milk sugar) 0.2 g;

sodium stearyl fumarate 0.005 g;

colloidal silicon dioxide (aerosil) 0.0025 g.

Compositions 1 and 2 are the closest analogues of the proposed technical solution.

Also known are preparations based on oxyethylammonium methylphenoxyacetate, having registration numbers of the Ministry of Health of the Russian Federation LP-005865, LP-005402, having the composition for 1 tablet:

hydroxyethylammonium methylphenoxyacetate 0.2 g;

Excipients:

potato starch 0.095 g;

lactose monohydrate (milk sugar) 0.2 g;

calcium stearate 0.005 g.

All noted compositions of hydroxyethylammonium methylphenoxyacetate are adaptogenic agents, but do not have antiviral activity.

The limited choice of drugs and the increase in drug resistance of viruses necessitate the development of new effective and safe drugs for the treatment of influenza. The authors of the proposed technical solution have made the assumption that a combination drug can be developed based on hydroxyethylammonium methylphenoxyacetate, which will exhibit antiviral activity against influenza A, antiviral properties against influenza B viruses, parainfluenza and seasonal coronavirus (for example, OS43).

The prerequisites for the development of the proposed composition were information about attempts to create forms of oxyethylammonium derivatives of methylphenoxyacetate (patent RU2394018, priority date: 07/25/2008) and their use as a means of activating alpha- and gamma-interferonogenesis (application EA200900805, priority date: 06/08. 2009), increasing the cytokine activity of total tryptophanyl-tRNA synthetase (patent RU2457837, priority date: 07/15/2011).

Thus, the technical problem, the solution of which is provided by the implementation or use of the proposed technical solution, and which could not be solved by the implementation or use of analogues, is the need to expand the arsenal of pharmaceutical compositions of antiviral drugs based on oxyethylammonium methylphenoxyacetate in solid dosage forms.

Disclosure of the invention

The technical problem is solved by using additional agents for the production of the drug hydroxyethylammonium methylphenoxyacetate, in combination with which the drug being created does not lose adaptogenic and immunomodulatory properties and acquires an antiviral effect, as well as by developing a stable solid pharmaceutical composition of hydroxyethylammonium methylphenoxyacetate with additional agents.

The use of hydroxyethylammonium methylphenoxyacetate in combination with o-cresol and/or 2-phenoxyacetic acid and/or 2-(3-methylphenoxy) acetic acid and/or 2-(4-methylphenoxy) acetic acid or a mixture of these substances and calcium and/or phosphorus and/or zinc or a mixture of elements for the production of a medicinal product with adaptogenic, immunomodulatory and antiviral activity against influenza A, influenza B, parainfluenza and coronavirus viruses.

Also proposed is a solid pharmaceutical composition with adaptogenic, immunomodulatory and antiviral properties, containing oxyethylammonium methylphenoxyacetate, additional components and excipients in the following ratio of components, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60

o-cresol 0.00001-0.5 and/or 2-phenoxyacetic acid and/or 2-(3-methylphenoxy) acetic acid and/or 2-(4-methylphenoxy) acetic acid or a mixture of these substances 0.00001-0 ,5

calcium 0.00001-0.75 and/or phosphorus 0.00001-0.29 and/or zinc 0.0000001-0.00050 or a mixture thereof

excipients up to 100.

The solid pharmaceutical composition is made in the dosage form of tablets or capsules.

Such a solid pharmaceutical composition can be prepared from the substance oxyethylammonium methylphenoxyacetate having a crystalline structure.

The proposed solid pharmaceutical composition can be used in conjunction with etiotropic therapy drugs, i.e. aimed at eliminating the cause of the disease.

A particular embodiment of the proposed solid pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 o-cresol 0.00001-0.2, and/or 2-phenoxyacetic acid, and/or 2-(3-methylphenoxy) acetic acid acid, and/or 2-(4-methylphenoxy) acetic acid acid, or a mixture of these substances 0.00001-0.2 Calcium 0.066–0.09, and/or Phosphorus 0.0017-0.014, and/or Zinc 0.0000085-0.00022, or a mixture thereof Excipients until 100 wt.% is obtained

A particular embodiment of the pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 Calcium 0.066–0.09, and/or Phosphorus 0.0017-0.014, and/or Zinc 0.0000085-0.00022, or a mixture thereof Excipients until 100 wt.% is obtained

A particular embodiment of the pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 o-cresol 0.00001-0.2 Calcium 0.066–0.09 Excipients until 100 wt.% is obtained

A particular embodiment of the pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 Calcium 0.066-0.09 Phosphorus 0.0017-0.014 Excipients until 100 wt.% is obtained

A particular embodiment of the pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 o-cresol 0.0001-0.2 Calcium 0.066-0.09 Phosphorus 0.0017-0.014 Excipients until 100 wt.% is obtained

A particular embodiment of the pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 o-cresol 0.0001-0.2, and/or 2-phenoxyacetic acid, and/or 2-(3-methylphenoxy) acetic acid, and/or 2-(4-methylphenoxy) acetic acid, or a mixture of these substances 0.00001-0.2 Calcium 0.066-0.09 Phosphorus 0.0017-0.014 Excipients until 100 wt.% is obtained

A particular embodiment of the pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 o-cresol 0.00001-0.2, and/or 2-phenoxyacetic acid, and/or 2-(3-methylphenoxy) acetic acid, and/or 2-(4-methylphenoxy) acetic acid, or a mixture of these substances 0.00001-0.2 Phosphorus 0.0017-0.014 Excipients until 100 wt.% is obtained

A particular embodiment of the pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 Phosphorus 0.0017-0.014 Excipients until 100 wt.% is obtained

A particular embodiment of the pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 o-cresol 0.00001-0.2 Phosphorus 0.0017-0.014 Excipients until 100 wt.% is obtained

A particular embodiment of the pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 o-cresol 0.00001-0.2, and/or 2-phenoxyacetic acid, and/or 2-(3-methylphenoxy) acetic acid, and/or 2-(4-methylphenoxy) acetic acid, or a mixture of these substances 0.00001-0.2 Zinc 0.0000085-0.00022 Excipients until 100 wt.% is obtained

A particular embodiment of the pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 Zinc 0.0000085-0.00022 Excipients until 100 wt.% is obtained

A particular embodiment of the pharmaceutical composition is the composition of the following composition, wt. %:

hydroxyethylammonium methylphenoxyacetate 20-60 o-cresol 0.00001-0.2 Zinc 0.0000085-0.00022 Excipients until 100 wt.% is obtained

The proposed compositions of oxyethylammonium methylphenoxyacetate differ from known compositions in that the composition of the proposed composition and its variants, in addition to the active substance oxyethylammonium methylphenoxyacetate, includes o-cresol from 0.00001 to 0.5%, 2-phenoxyacetic acid, 2-(3-methylphenoxy) acetic acid acid and 2-(4-methylphenoxy) acetic acid from 0.00001 to 0.5% or a mixture thereof, based on the weight of the active substance. In addition, the composition contains calcium in a relative amount from 0.00001 mg to 0.75 mg based on the total weight of the tablet (0.5 g), phosphorus in a relative amount from 0.00001 mg to 0.29 mg based on the total tablet weight (0.5 g), zinc in a relative amount from 0.0000001 mg to 0.00050 mg based on the total tablet weight (0.5 g).

In this case, the composition will not necessarily contain all components; it is possible to contain only o-cresol or 2-phenoxyacetic acid, or 2-(3-methylphenoxy) acetic acid, or 2-(4-methylphenoxy) acetic acid, or a mixture thereof, or calcium, or phosphorus, or zinc, or combinations thereof.

The technical result of the proposed technical solution is to obtain the drug oxyethylammonium methylphenoxyacetate with increased adaptogenic, immunomodulatory and antiviral effects. The technical result of the proposed composition of hydroxyethylammonium methylphenoxyacetate is the manifestation of antiviral properties, increased bioavailability, and reduction of negative side reactions in the claimed compositions, achieved through the use of additional components.

The adaptogenic effect of the proposed technical solution is determined by the presence in the composition of oxyethylammonium methylphenoxyacetate (tris(2-hydroxyethyl)ammonium 2-methylphenoxyacetate; tris(2-oxyethyl)ammonium orthocresoxyacetate) - a synthetic adaptogen that leads a living organism (including humans) to a state of nonspecifically increased resistance to various diseases and unfavorable environmental conditions (adaptation to temperature changes and atmospheric pressure, lack of oxygen, ionizing radiation, carcinogenic agents, industrial pollution, infections, etc.). Adaptogens are harmless to the body and have a wide range of stimulating physiological effects.

The adaptogenic effect of oxyethylammonium methylphenoxyacetate is known from the state of the art (Shabanov P.D., Mokrenko E.V. New immunomodulator and adaptogen trekrezan as a means of prevention and treatment of inflammatory colds // Bulletin of the Smolensk State Medical Academy. 2014. No. 2. URL: https: //cyberleninka.ru/article/n/novyy-immunomodulyator-i-adaptogen-trekrezan-kak-sredstvo-profilaktiki-i-lecheniya-prostudnyh-vospalitelnyh-zabolevaniy (date of access: 01/26/2023).

The adaptogenicity of oxyethylammonium methylphenoxyacetate is manifested in stimulating the production of alpha and gamma interferons, influencing the immune status of the body due to the activation of cellular and humoral immunity, stimulating the phagocytic activity of macrophages, strengthening the body's immune system, increasing endurance during physical and mental stress, reducing the effect of various toxins, increasing the body's resistance to hypoxia, low and high temperatures and other unfavorable environmental factors. These results were confirmed by the results of studies conducted by both the applicant and other researchers and are known from the state of the art (Shabanov P.D., Zarubina I.V., Mokrenko E.V. Pharmacology of trekrezan - a new immunomodulator and adaptogen // Clinical Reviews. pharmacocol. and medicinal therapy. 2014. No. 2. URL: https://cyberleninka.ru/article/n/farmakologiya-trekrezana-novogo-immunomodulyatora-i-adaptogena (date of access: 06.29.2021); Dyakov V. M. ., Kazimirovskaya V.B., Kovalchuk S.F.M., 2005. URL: http://content.medsovet.info/files2/forum/125479/Trekrezan.pdf (access date: 01/26/2023).

When mice were administered a placebo - 0.1 ml aseptic saline solution (control group) - minimal concentrations of interferons were detected in the serum of the animals. On average, no more than 2.0 IU/ml of α- and 2.4 IU/ml of γ-interferon were detected within 24 hours after intraperitoneal administration. Intraperitoneal administration of oxyethylammonium methylphenoxyacetate 25 mg/kg to mice already after 6 hours led to a significant (p <0.05) increase in interferon levels in animals compared to the placebo group. During this time, 54.2 IU/ml interferon, resistant to pH and heating (α-type), was formed. γ-Interferon, calculated from the difference between total and α-interferon, was within 14.4 IU/ml and its content did not differ significantly from the placebo group. A day later this ratio changed. α-Interferon on average was minimal - 8.3 IU/ml, and γ-interferon increased significantly to 43.2 IU/ml (p <0.05).

Polynucleotide I:C (positive control) under similar conditions induced only α-interferon, which after 6 hours reached a level of 160.6 IU/ml and was significantly higher (p metaprot ≈ oxyethylammonium methylphenoxyacetate). In addition, oxyethylammonium methylphenoxyacetate, used as monotherapy and in combination with other immunomodulators, increases the lymphokine-producing function of lymphocytes, the phagocytic activity of neutrophils, the activity of oxygen-independent microbicidal systems of phagocytes, reduces the phagocytic number, the rate of completion of phagocytosis and the activity of oxygen-dependent microbicidal systems of phagocytes in acute bronchopulmonary inflammation rats

In experiments performed in vitro on alveolar and peritoneal macrophages to evaluate the antiradical (antisuperoxide) effect of immunomodulators, it was shown that the effect of oxyethylammonium methylphenoxyacetate, metaprote, polyoxidonium and their combinations on lucigenin-dependent biochemiluminescence was also different and dose-dependent. In these systems, immunomodulators at different concentrations had both pro-oxidant and antioxidant effects, while hydroxyethylammonium methylphenoxyacetate exhibited an antioxidant effect.

Thus, oxyethylammonium methylphenoxyacetate should be considered as a highly effective adaptogen, an immunomodulator that activates all forms of immunity (cellular, humoral, phagocytosis). The simple chemical structure, low toxicity, and low cost of hydroxyethylammonium methylphenoxyacetate allow the drug to be used as a drug of choice when prescribing immunomodulators. It should be emphasized that hydroxyethylammonium methylphenoxyacetate has both a direct immunostimulating effect and a powerful antiasthenic effect, which helps reduce the manifestations of inflammation and toxicosis in viral colds. These properties of oxyethylammonium methylphenoxyacetate make it a unique drug in the sense of both the prevention of acute respiratory viral infections due to its pronounced adaptogenic properties, and the treatment of colds, which is due to the anti-inflammatory, reparative and ergotropic properties of oxyethylammonium methylphenoxyacetate.

The use of oxymethylammonium methylpheninoxyacetate in combination with additional agents leads to the normalization of metabolic processes, which in turn leads to an increase in the body's resistance to influenza and ARVI, as well as an increase in the body's resistance to stressful situations and adverse environmental influences.

It has been revealed that the introduction of certain doses of calcium and/or phosphorus and/or zinc into compositions containing methylphenoxyacetate as the active ingredient significantly increases the therapeutic effectiveness of the composition, namely, the developed composition exhibits not only adaptogenic and immunomodulatory effects, but also antiviral ones.

This effect of the combination of calcium and/or phosphorus and/or zinc or a mixture thereof and oxyethylammonium methylphenoxyacetate is unexpected, although the role of each element in the human body is known.

Without calcium and phosphorus, it is impossible to launch biochemical processes - the formation of enzymes (specific proteins, enzymes, amino acids) that contribute to normal phosphorus-calcium metabolism.

Calcium is the second element involved in various physiological and biochemical processes occurring in the body. It plays an important role in regulating the permeability of cell membranes, in the molecular mechanism of muscle contraction, digestive and endocrine glands, in activating various enzymatic systems, including those responsible for blood clotting, etc. In the systemic regulation of calcium homeostasis, an important role is played by calcium-dependent receptors, which are isolated in the parathyroid glands (inhibit the secretion of PTH), vascular smooth muscle cells, cardiomyocytes, vascular endothelial cells, as well as in the parathyroid glands, kidneys, and intestines. One of the important effects of calcium-dependent receptors is the regulation of vascular tone and blood pressure (BP), which is carried out through modulation of calcium homeostasis.

Phosphorus is one of the vital microelements that takes part in metabolic processes. It is part of all cells of the body. Phosphates maintain acid-base balance at a physiological level and contribute to the production of cellular energy for metabolic reactions.

Zinc is one of the essential microelements - these are microelements without which the body cannot grow, develop and complete its natural life cycle. Its main function is that it is a cofactor for enzymes (LDH, carbonic anhydrase, RNA and DNA polymerases).

The maximum daily dose for an adult: calcium is 2500 mg, phosphorus 1600 mg, zinc 25 mg. Therefore, when additionally introducing calcium, phosphorus and zinc salts, you should not exceed the daily dose.

As will be shown in the studies below, the independent use of additional components: o-cresol and/or 2-phenoxyacetic acid and/or 2-(3-methylphenoxy) acetic acid and/or 2-(4-methylphenoxy) acetic acid and/or calcium and/or phosphorus, and/or zinc or a mixture thereof does not lead to a reliably significant effect at the indicated concentrations. Calcium, phosphorus, and zinc do not have independent therapeutic efficacy against antiviral diseases. It is a mistake to think that by using one microelement you can strengthen the immune system, since immunity is a complex of various reactions in the body - cellular, humoral, etc.

As part of the proposed technical solution, calcium and/or phosphorus and/or zinc or a mixture thereof lead to increased bioavailability and increased adaptogenic, immunostimulating and antiviral properties of the drug hydroxyethylammonium methylphenoxyacetate and significantly increase the effectiveness of the latter due to the use of the drug in the composition.

In the production of the proposed composition, calcium and zinc are used in the form of chelates, stearates, phosphorus in the form of orthophosphoric acid and its salts.

The authors of this technical solution conducted a comparative study of the antiviral activity of Trecrezan preparations without the addition of compositions (1) and (2), as well as compositions (1) and (2) containing different combinations of additives No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc.

Compositions for the study: composition 1 :

hydroxyethylammonium methylphenoxyacetate 0.2 g;

Excipients:

potato starch 0.0925 g;

lactose monohydrate (milk sugar) 0.2 g;

sodium stearyl fumarate 0.005 g;

colloidal silicon dioxide (aerosil) 0.0025 g.

With remainder 2 :

hydroxyethylammonium methylphenoxyacetate 0.2 g;

Excipients:

mycrystalline cellulose 0.0925 g;

lactose monohydrate (milk sugar) 0.2 g;

sodium stearyl fumarate 0.005 g;

colloidal silicon dioxide (aerosil) 0.0025 g.

To study compositions (1) and (2), containing different combinations of additives, the following compositions were used: composition-additive (wt.%) :

hydroxyethylammonium methylphenoxyacetate 40;

additional components - according to research

excipients up to 100.

As auxiliary components we took - in the variant, composition 1-add: potato starch, lactose monohydrate, sodium stearyl fumarate, colloidal silicon dioxide; in the version 2-additional composition: microcrystalline cellulose, lactose monohydrate, sodium stearyl fumarate, colloidal silicon dioxide.

The purpose of the study was a comparative assessment of the antiviral activity of the drug Trekrezan without the addition of compositions (1) and (2), as well as compositions (1) and (2) containing different combinations of additives No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc in relation to influenza A and B viruses, parainfluenza and seasonal coronavirus OS43 in vitro . The objectives of the study included:

1.In vitro determination of the dose of the drug at which 50% of the population of the studied cells dies(Further TsTD ₅₀ ) for three cell cultures (first) - cocker spaniel kidneys(hereinafter MDCK), (second) - epidermoid carcinoma of the human larynx(Further HEr-2)and (thirds) - renal epithelium of the green monkey(Further Vero).

2.In vitro determination of the antiviral activity of the study drug againstinfluenza A viruses (strain A/Aichi/2/68 (H3N2)) and influenza B (strain B/Malaysia/2506/04) on cell cultureMDCKmethod hemagglutination reactions(hereinafter referred to as RGA) by chemotherapy index(hereinafter referred to as HTI or TsTD ₅₀ /ED ₅₀ , where ED₅₀ is the 50% effective dose of the study drug).

3. In vitro determination of the antiviral activity of the studied drugs against parainfluenza virus type 3 (strain III v2932) on HEp-2 cell culture using enzyme-linked immunosorbent assay (ELISA) according to the chemotherapy index (hereinafter referred to as CTI or CTD ₅₀ /ED ₅₀ , where ED ₅₀ is the 50% effective dose of the study drug).

4. In vitro determination of the antiviral activity of the study drugs against the seasonal coronavirus OS43 on Vero cell culture using the microtetrazolium test (MTT test) according to the chemotherapy index (hereinafter referred to as CTI or CTD ₅₀ /ED ₅₀ , where ED ₅₀ is the 50% effective dose of the test drug drug).

During the study, it was found that the proposed composition (with additional components) exhibited increased antiviral activity against all viruses used, most pronounced against the influenza A virus .

Influenza viruses A/Aichi/2/68(H3N2) and B/Malaysia/2506/04 were cultured in developing chicken embryos. Next, a series of aliquots were prepared from the virus-containing allantoic fluid. The resulting aliquots were stored at -80°C.

Human parainfluenza virus type 3, strain III v2932 , was accumulated on a permissive culture of HEp-2 cells, after which a series of aliquots were prepared from the culture liquid. The resulting aliquots were stored at -80°C.

Seasonal human coronavirus OS43 was accumulated on a permissive Vero cell culture, after which a series of aliquots were prepared from the culture liquid. The resulting aliquots were stored at -80°C.

The cell suspension (2-3*10 ⁶ cells/ml) was scattered into 96-well microplates at 100 μl/well and cultured for 24 hours at 37°C in an atmosphere of 5% CO ₂ until a confluent monolayer was achieved. The integrity of the cell monolayer was assessed using an inverted microscope.

The results of determining the 50% cytotoxic dose of the drug Trecrezan composition (1) and (2) without additives (CTD ₅₀ ) in relation to all three cell cultures used in the work are presented in average form in Table 1.

Table 1 - Average results of determining the 50% cytotoxic dose of the drug Trecrezan (compositions (1) and (2) without additives) CTD ₅₀ on MDCK, Vero and HEp-2 cell cultures Cell culture Concentration, µg/ml % cell viability CTD ₅₀ , µg/ml MDCK
(cocker spaniel kidney cells) 4000 64.35±5.53 4250 2000 91.06±5.37 1000 95.86±14.83 500 85.33±8.13 250 83.53±6.25 125 79.29±2.25 64 81.81±0.10 0 100 NEr-2
(human epidermoid carcinoma of the larynx) 4000 17.77±0.40 1165 2000 44.94±7.64 1000 59.41±2.93 500 67.03±3.92 250 81.73±0.57 125 88.30±1.39 64 92.00±5.19 0 100 Vero
(renal epithelium of green monkey) 4000 13.24±4.77 1374 2000 48.71±2.80 1000 69.02 0.02 500 78.25±1.94 250 81.64±3.08 125 81.30±0.53 64 84.09±0.59 0 100

From the data presented in Table 1 , it is clear that the sensitivity of cell lines to the action of the drug is generally similar for the Vero and HEp-2 lines, while for the MDCK cell line it is less pronounced.

The CTD ₅₀ value for Trecrezan compositions (1) and (2) without additives for the MDCK cell line exceeded 4000 μg/ml (calculated value 4250 μg/ml), therefore, a dose of 3000 μg/ml was used as the first working concentration for experiments with influenza viruses ml.

On the HEp-2 and Vero cell lines, the CTD ₅₀ values were 1165 and 1374 μg/ml, respectively. At the same time, at dosages of 1000 and 500 μg/ml, which are formally below the CTD ₅₀ , residual cytotoxicity was observed, which led to viability values in the range of 59-78%. In connection with the noted effect of a gradual decrease in the cytotoxicity of the drug, depending on the dosage, it was decided to use a dose of 300 μg/ml as the first concentration when working with the parainfluenza virus and seasonal coronavirus .

Additionally, experiments were carried out with the drug Trekrezan of compositions (1) and (2) with the introduction of different concentrations and sets of additives, No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid , No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc. The results of determining the cytotoxicity of the drug Trekrezan (composition (1) and (2) with additives) in relation to all three cell cultures used in the work are presented in average form in Tables 1.1-1.3.

Table 1.1 - Average results of determining the 50% cytotoxic dose of the drug Trekrezan (compositions (1) and (2) with additives) CTD ₅₀ on MDCK cell culture CTD ₅₀ , µg/ml Serial numbers of added additives No. 1 No. 2 No. 3 No. 4 No. 5 No. 6 No. 7 The concentration of the additive is indicated as a percentage of the active substance contained in the tablet The concentration of the additive is indicated in (mg)
per tablet weight 4438 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 4419 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 4414 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 4410 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 4406 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 4397 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 4395 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 4391 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 4387 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 4386 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 4382 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 4381 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 4378 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 4378 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 4373 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 4369 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 4367 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 4365 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 4363 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 4363 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 4359 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 4357 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 4354 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 4354 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 4353 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 4350 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 4349 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 4346 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 4345 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 4341 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 4340 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 4339 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 4337 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 4335 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 4335 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 4331 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 4329 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 4326 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 4325 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 4325 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 4322 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 4322 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 4321 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 4318 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 4316 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 4313 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 4312 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 4311 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 4308 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 4307 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 4306 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 4303 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 4301 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 4298 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 4297 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 4297 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 4294 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 4294 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 4293 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 4290 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 4288 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 4284 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 4284 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 4282 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 4280 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 4279 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 4278 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 4274 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 4273 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 4270 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 4269 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 4269 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 4266 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 4265 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 4265 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 4262 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 4260 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 4256 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 4256 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 4254 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 4252 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 4250 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 4250 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 4246 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 4245 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 4241 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 4241 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 4241 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 4238 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 4237 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 4237 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 4233 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 4232 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 4228 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 4228 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 4226 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 4224 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 4222 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 4222 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 4218 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 4217 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 4213 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 4213 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 4209 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 4209 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 4209 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 4205 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 4204 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 4200 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 4200 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 4198 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 4196 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 4194 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 4190 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 4185 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 4185 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 4181 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 4181 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 4177 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 4176 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 4172 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 4168 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 4166 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 4157 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 4153 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 4149 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 4144 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 4125 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000

Conclusion: as can be seen from the data presented in Table 1.1 , the MDCK cell line exhibits the least sensitivity to Trecrezan of compositions (1) and (2) containing a minimum amount of additives , No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid and the maximum amount of additives , No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc.

Table 1.2 - Average results of determining the 50% cytotoxic dose of the drug Trekrezan (compositions (1) and (2) with additives) CTD ₅₀ on HEp-2 cell culture CTD ₅₀ , µg/ml Serial numbers of added additives No. 1 No. 2 No. 3 No. 4 No. 5 No. 6 No. 7 The concentration of the additive is indicated as a percentage of the active substance contained in the tablet The concentration of the additive is indicated in (mg) per tablet weight 1179 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 1178 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 1175 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 1173 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 1173 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 1171 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 1171 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 1171 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 1170 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 1170 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 1169 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 1169 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 1168 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 1168 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 1167 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 1167 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 1166 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 1166 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 1166 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 1166 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 1165 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 1165 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 1165 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 1164 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 1164 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 1164 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 1163 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 1163 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 1163 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 1163 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 1162 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 1162 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 1162 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 1162 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 1161 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 1161 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 1161 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 1160 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 1160 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 1160 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 1160 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 1160 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 1159 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 1159 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 1159 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 1159 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 1159 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 1158 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 1158 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 1158 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 1158 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 1158 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 1158 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 1157 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 1157 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 1157 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 1156 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 1156 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 1156 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 1156 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 1156 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 1155 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 1155 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 1155 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 1155 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 1155 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 1155 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 1154 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 1154 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 1154 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 1154 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 1153 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 1153 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 1153 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 1152 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 1152 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 1152 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 1152 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 1152 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 1151 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 1151 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 1151 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 1151 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 1151 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 1151 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 1150 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 1150 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 1150 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 1150 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 1150 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 1149 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 1149 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 1149 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 1149 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 1148 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 1148 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 1148 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 1147 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 1147 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 1147 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 1147 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 1146 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 1146 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 1146 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 1146 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 1145 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 1145 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 1145 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 1144 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 1144 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 1143 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 1143 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 1143 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 1143 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 1142 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 1142 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 1141 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 1141 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 1140 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 1140 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 1139 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 1139 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 1138 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 1137 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 1136 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 1135 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 1132 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 1131 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000

Conclusion: as can be seen from the data presented in Table 1.2 , the HEp-2 cell line, like the previously tested MDCK , exhibits the least sensitivity to Trecrezan of compositions (1) and (2) containing a minimum amount of additives , No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid and the maximum amount of additives , No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc.

Table 1.3 - Average results of determining the 50% cytotoxic dose of the drug Trekrezan (compositions (1) and (2) with additives) CTD ₅₀ on Vero cell culture CTD ₅₀ , µg/ml Serial numbers of added additives No. 1 No. 2 No. 3 No. 4 No. 5 No. 6 No. 7 The concentration of the additive is indicated as a percentage of the active substance contained in the tablet The concentration of the additive is indicated in (mg) per tablet weight 1394 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 1392 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 1388 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 1386 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 1386 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 1385 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 1384 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 1383 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 1383 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 1382 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 1381 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 1380 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 1380 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 1380 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 1379 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 1378 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 1378 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 1377 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 1377 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 1377 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 1376 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 1375 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 1375 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 1375 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 1374 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 1374 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 1374 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 1374 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 1374 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 1373 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 1373 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 1373 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 1372 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 1372 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 1371 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 1371 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 1371 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 1371 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 1371 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 1370 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 1370 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 1370 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 1369 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 1369 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 1369 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 1368 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 1368 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 1368 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 1368 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 1367 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 1367 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 1367 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 1366 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 1366 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 1366 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 1366 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 1365 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 1365 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 1365 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 1365 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 1364 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 1364 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 1364 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 1364 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 1364 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 1363 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 1363 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 1363 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 1362 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 1362 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 1362 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 1362 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 1362 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 1361 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 1361 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 1361 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 1361 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 1360 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 1360 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 1360 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 1359 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 1359 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 1359 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 1359 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 1358 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 1358 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 1358 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 1358 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 1357 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 1357 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 1356 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 1356 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 1356 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 1356 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 1355 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 1355 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 1355 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 1355 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 1354 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 1354 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 1353 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 1353 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 1353 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 1353 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 1352 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 1352 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 1352 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 1351 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 1350 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 1350 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 1350 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 1349 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 1349 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 1349 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 1347 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 1347 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 1347 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 1346 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 1345 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 1345 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 1344 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 1343 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 1343 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 1341 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 1341 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 1339 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 1336 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 1334 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000

Conclusions: as can be seen from the data presented in Table 1.3 , the Vero cell line, like the previously tested HEp-2 and MDCK , exhibits the least sensitivity to Trecrezan of compositions (1) and (2) containing a minimum amount of additives , No. 1 - o-cresol , No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid and the maximum amount of additives , No. 5 - calcium, No. 6 - phosphorus and No. 7 – zinc.

The results of a study of the antiviral activity of the drug Trekrezan (compositions (1) and (2) without additives ) against influenza A and B viruses are presented in average form in Table 2.

Table 2 - Average results of testing the antiviral activity of the drug Trekrezan (compositions (1) and (2) without additives ) against influenza A and B viruses in vitro on MDCK cell culture Virus Concentration, µg/ml Virus titer, lg TID ₅₀ Decrease in titer compared to control Decrease in infectious activity compared to control, % ED ₅₀ A/Aichi/2/68 (H3N2) 3000 4.83±0.29 2 99 67.1 1000 5.33±0.29 1.5 96.84 333 6.33±0.29 0.5 68.38 111 6.33±0.29 0.5 68.38 33 6.67±0.29 0.17 31.87 0 6.83±0.29 - - B/Malaysia/2506/04 3000 4.33±0.29 1.67 97.85 243.7 1000 4.83±0.76 1.17 93.19 333 5.67±0.29 0.33 53.58 111 6.0±1.0 0.00 0.00 33 5.83±1.15 0.17 31.87 0 6.0±0.87 - -

As follows from the data presented in Table 2, it is clear that the average ED values of ₅₀ Trecrezan preparations of composition (1) and (2) without additives for the influenza A virus was 64.4. This leads to the value of the chemotherapeutic index CTI = 4250/67.1 = 63.3, which significantly exceeds the threshold value of 8 and indicates the presence of antiviral activity of this drug. At the same time, for the influenza B virus, the antiviral activity was less pronounced, although the CTI value was 4250/243.7 = 17.4, which also exceeds the threshold value.

Additionally, experiments were carried out with the drug Trekrezan of compositions (1) and (2) with the introduction of different concentrations and sets of additives, No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid , No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc. The results of determining the antiviral activity according to ED ₅₀ (50% effective dose) of the drug Trekrezan (compositions (1) and (2) with additives ) are presented in average form in tables 2.1-2.4. The average results for two compositions are presented.

Table 2.1 - Average results of determining the antiviral activity according to ED ₅₀ (50% effective dose) of the drug Trecrezan (compositions (1) and (2) with additives ) against the influenza A virus (Aichi/2/68 (H3N2)) in vitro in culture MDCK cells ED ₅₀ , mcg/ml Serial numbers of added additives No. 1 No. 2 No. 3 No. 4 No. 5 No. 6 No. 7 The concentration of the additive is indicated as a percentage of the active substance contained in the tablet The concentration of the additive is indicated in (mg)
per tablet weight 10.3 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 11.4 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 12.5 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 13.7 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 13.7 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 14.8 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 14.8 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 15.9 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 15.9 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 17.0 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 17.0 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 18.1 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 18.1 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 19.3 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 19.4 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 20.4 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 20.5 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 21.5 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 21.7 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 22.8 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 22.8 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 23.9 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 23.9 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 24.0 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 25.0 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 25.0 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 25.1 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 26.1 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 26.1 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 26.2 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 27.3 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 27.3 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 27.3 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 27.3 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 28.4 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 28.5 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 28.5 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 29.5 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 29.6 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 29.6 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 30.6 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 30.7 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 30.7 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 31.8 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 31.8 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 32.9 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 33.1 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 33.1 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 34.1 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 34.2 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 34.2 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 35.2 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 35.3 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 35.3 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 36.5 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 36.5 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 36.5 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 36.5 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 37.6 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 37.6 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 37.6 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 37.6 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 38.7 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 38.7 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 38.7 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 38.7 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 39.8 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 39.8 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 39.8 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 39.8 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 40.9 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 40.9 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 40.9 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 40.9 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 42.1 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 42.1 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 42.2 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 43.2 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 43.2 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 43.3 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 44.3 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 44.3 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 44.5 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 45.6 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 45.6 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 46.7 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 46.7 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 46.8 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 47.8 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 47.8 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 47.9 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 48.9 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 48.9 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 49.0 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 50.1 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 50.1 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 50.1 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 50.1 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 51.2 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 51.3 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 51.3 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 52.3 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 52.4 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 52.4 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 53.4 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 53.5 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 53.5 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 54.6 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 54.6 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 55.7 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 55.9 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 56.9 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 57.0 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 58.0 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 58.1 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 59.3 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 59.3 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 60.4 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 60.4 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 61.5 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 61.5 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 62.6 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 62.6 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 63.7 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 63.7 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 64.9 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 66.0 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 67.1 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000

Conclusions: as can be seen from the data presented in Table 2.1 , the greatest antiviral activity against influenza A virus (Aichi/2/68 (H3N2)) in vitro on MDCK cell culture is exhibited by the drug Trekrezan of compositions (1) and (2), containing the maximum number of additives , No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc, while the introduction of any of the tested additives or their set into the Trekrezan preparation, both in the case of composition (1) and in the case of composition (2), leads to an increase in antiviral activity, which is expressed in a decrease in detectable 50 % effective concentration.

Table 2.2 - Average results of assessing the antiviral activity according to the chemotherapeutic index (CHI) of the drug Trekrezan (compositions (1) and (2) with additives) against the influenza A virus (Aichi/2/68 (H3N2)) in vitro on MDCK cell culture

HTI = CTD ₅₀ /ED ₅₀ Serial numbers of added additives No. 1 No. 2 No. 3 No. 4 No. 5 No. 6 No. 7 The concentration of the additive is indicated as a percentage of the active substance contained in the tablet The concentration of the additive is indicated in (mg)
per tablet weight 424.2 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 384.0 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 351.3 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 320.9 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 319.8 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 296.4 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 295.5 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 277.4 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 275.3 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 259.9 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 258.5 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 243.4 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 242.1 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 227.3 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 223.7 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 216.4 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 211.8 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 206.4 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 200.6 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 191.1 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 190.6 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 182.0 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 181.6 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 177.3 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 174.5 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 173.8 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 169.6 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 167.2 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 166.8 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 162.9 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 159.7 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 159.3 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 156.4 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 156.1 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 153.3 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 149.6 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 149.3 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 147.9 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 144.4 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 143.9 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 142.7 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 139.4 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 139.1 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 134.4 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 134.1 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 130.2 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 129.7 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 127.7 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 126.1 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 125.5 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 123.6 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 122.5 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 121.6 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 120.1 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 118.5 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 118.3 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 116.2 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 116.0 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 114.9 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 114.7 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 112.7 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 112.4 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 111.9 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 111.5 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 109.7 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 109.4 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 108.9 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 108.7 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 106.8 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 106.6 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 105.8 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 105.6 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 103.8 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 103.6 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 102.7 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 101.5 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 100.7 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 100.3 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 98.9 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 98.4 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 97.9 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 96.6 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 96.0 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 94.3 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 94.1 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 91.9 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 91.7 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 90.0 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 89.7 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 89.7 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 88.1 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 87.9 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 87.7 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 85.9 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 85.8 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 85.7 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 84.1 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 83.9 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 83.9 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 82.3 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 82.0 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 81.8 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 80.7 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 80.5 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 80.2 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 78.9 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 78.7 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 77.2 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 77.0 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 75.6 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 74.2 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 73.8 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 72.8 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 72.8 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 71.9 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 70.5 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 70.1 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 69.0 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 68.6 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 68.0 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 67.5 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 66.9 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 66.7 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 65.6 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 65.4 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 64.3 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 63.4 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 63.3 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000

Conclusions: as can be seen from the data presented in table 2.2 , all obtained CTI values significantly exceed the threshold value of 8, which indicates the presence of antiviral activity of Trekrezan compositions (1) and (2) with any combination of additives, while the greatest antiviral activity against influenza A virus (Aichi/2/68 (H3N2)) in vitro on MDCK cell culture exhibits Trecrezan of both composition (1) and (2) containing the maximum amount of additives , No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid , No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc, while introducing any of the tested additives or their inclusion in the drug Trekrezan, both in the case of composition (1) and in the case of composition (2), leads to an increase in antiviral activity, which is expressed in an increase in the estimated CTI index.

Table 2.3 - Average results of determining the antiviral activity according to ED ₅₀ (50% effective dose) of the drug Trecrezan (compositions (1) and (2) with additives) against the influenza B virus (Malaysia/2506/04) in vitro on MDCK cell culture ED ₅₀ , mcg/ml Serial numbers of added additives No. 1 No. 2 No. 3 No. 4 No. 5 No. 6 No. 7 The concentration of the additive is indicated as a percentage of the active substance contained in the tablet The concentration of the additive is indicated in (mg)
per tablet weight 37.4 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 41.5 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 45.5 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 49.6 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 49.6 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 53.7 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 53.7 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 57.7 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 57.7 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 61.8 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 61.8 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 65.9 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 65.9 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 70.0 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 70.5 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 74.0 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 74.6 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 78.1 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 78.7 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 82.7 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 82.7 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 86.8 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 86.8 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 87.1 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 90.9 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 90.9 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 91.2 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 94.9 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 94.9 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 95.2 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 99.0 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 99.0 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 99.3 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 99.3 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 103.1 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 103.4 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 103.4 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 107.1 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 107.4 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 107.4 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 111.2 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 111.5 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 111.5 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 115.6 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 115.6 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 119.6 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 120.2 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 120.2 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 123.7 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 124.3 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 124.3 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 127.8 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 128.4 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 128.4 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 132.4 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 132.4 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 132.4 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 132.4 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 136.5 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 136.5 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 136.5 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 136.5 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 140.6 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 140.6 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 140.6 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 140.6 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 144.6 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 144.6 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 144.6 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 144.6 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 148.7 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 148.7 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 148.7 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 148.7 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 152.8 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 152.8 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 153.3 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 156.8 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 156.8 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 157.4 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 160.9 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 160.9 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 161.5 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 165.5 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 165.5 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 169.6 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 169.6 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 169.9 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 173.7 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 173.7 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 174.0 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 177.7 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 177.7 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 178.0 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 181.8 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 181.8 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 182.1 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 182.1 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 185.9 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 186.2 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 186.2 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 189.9 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 190.2 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 190.2 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 194.0 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 194.3 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 194.3 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 198.4 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 198.4 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 202.4 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 203.0 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 206.5 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 207.1 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 210.6 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 211.2 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 215.2 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 215.2 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 219.3 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 219.3 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 223.4 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 223.4 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 227.4 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 227.4 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 231.5 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 231.5 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 235.6 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 239.6 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 243.7 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000

Conclusions: as can be seen from the data presented in Table 2.3 , Trekrezan, both composition (1) and composition (2) containing the maximum number of additives, No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc, while the introduction of any of the tested additives or their set into the Trekrezan preparation, both in the case of composition (1) and in the case of composition (2), leads to an increase in antiviral activity, which is expressed in a decrease in detectable 50 % effective concentration.

Table 2.4. − Average results of assessing the antiviral activity according to the chemotherapeutic index (CHI) of the drug Trekrezan (compositions (1) and (2) with additives) against the influenza B virus (Malaysia/2506/04) in vitro on MDCK cell culture HTI = CTD ₅₀ /ED ₅₀ Serial numbers of introduced additives No. 1 No. 2 No. 3 No. 4 No. 5 No. 6 No. 7 The concentration of the additive is indicated as a percentage of the active substance contained in the tablet The concentration of the additive is indicated in (mg) per tablet weight 115.3 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 104.6 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 95.3 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 88.0 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 87.6 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 81.4 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 81.1 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 75.9 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 75.7 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 71.1 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 70.8 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 66.9 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 66.6 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 63.1 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 60.9 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 59.6 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 57.9 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 56.8 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 54.9 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 52.6 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 52.3 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 50.2 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 49.9 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 48.2 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 48.0 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 47.9 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 46.4 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 46.1 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 45.9 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 44.4 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 44.3 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 44.1 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 42.8 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 42.6 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 42.6 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 41.2 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 41.0 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 41.0 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 39.7 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 39.7 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 39.6 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 38.4 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 38.3 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 37.1 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 37.0 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 36.0 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 35.4 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 34.8 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 34.7 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 34.5 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 33.9 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 33.8 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 33.3 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 32.7 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 32.5 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 32.4 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 32.0 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 31.8 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 31.6 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 31.5 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 31.1 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 30.9 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 30.8 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 30.7 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 30.2 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 30.1 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 30.0 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 29.9 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 29.5 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 29.4 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 29.2 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 29.1 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 28.7 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 28.6 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 28.5 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 28.0 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 27.8 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 27.6 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 27.3 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 27.2 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 27.1 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 26.8 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 26.4 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 25.9 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 25.8 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 25.3 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 25.2 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 24.8 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 24.7 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 24.4 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 24.3 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 24.2 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 24.0 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 23.8 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 23.7 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 23.4 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 23.3 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 23.0 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 22.9 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 22.8 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 22.6 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 22.5 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 22.5 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 22.2 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 22.1 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 21.8 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 21.7 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 21.4 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 21.3 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 21.0 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 20.5 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 20.3 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 20.3 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 20.1 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 19.6 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 19.4 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 19.3 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 19.1 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 19.0 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 18.8 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 18.7 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 18.5 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 18.4 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 18.2 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 18.1 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 17.9 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 17.6 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 17.4 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000

Conclusions: as can be seen from the data presented in Table 2.4 , all obtained CTI values significantly exceed the threshold value, which indicates the presence of antiviral activity of Trecrezan compositions (1) and (2) with any combination of additives, while the greatest antiviral activity against the virus influenza B (Malaysia/2506/04) in vitro on MDCK cell culture exhibits Trecrezan of both compositions (1) and (2) containing the maximum amount of additives, No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc, while introducing any of the tested additives or a set of them into the preparation Trekrezan, both in the case of composition (1) and in the case of composition (2), leads to an increase in antiviral activity, which is expressed in an increase in the estimated CTI index .

The results of a study of the antiviral activity of Trecrezan drugs composition (1) and (2) without additives against human parainfluenza virus type 3 are presented in average form in Table 3.

Table 3 - Results of testing the antiviral activity of Trecrezan against parainfluenza virus type 3 in vitro Concentration, µg/ml Virus titer, lg TID ₅₀ Decrease in titer compared to control Decrease in infectious activity compared to control, % ED ₅₀ 300 1.83±0.29 1.17 93.19 26.5 100 2.33±0.29 0.67 78.46 33 2.67±0.29 0.33 53.58 eleven 3.17±0.58 - - 3.3 3.17±0.58 - - 0 3.0±0 - -

From the data presented in Table 3 it is clear that the ED value of ₅₀ Trecrezan preparations of composition (1) and (2) without additives for the parainfluenza virus was 26.5. This results in a value of HTI = 1165/26.5 = 44, which exceeds the threshold value of 8.

Additionally, experiments were carried out with the drug Trekrezan of compositions (1) and (2) with the introduction of different concentrations and sets of additives, No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid , No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc. The results of determining the antiviral activity according to ED ₅₀ (50% effective dose) of the drug Trekrezan (compositions (1) and (2) with additives) are presented in average form in Tables 3.1-3.2.

Table 3.1 - Average results of determining antiviral activity according to ED ₅₀ (50% effective dose) of the drug Trekrezan (compositions (1) and (2) with additives) against parainfluenza virus type 3 in vitro on HEp-2 cell culture. ED ₅₀ ,
µg/ml Serial numbers of added additives No. 1 No. 2 No. 3 No. 4 No. 5 No. 6 No. 7 The concentration of the additive is indicated as a percentage of the active substance contained in the tablet The concentration of the additive is indicated in (mg)
per tablet weight 4.1 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 4.5 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 5.0 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 5.4 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 5.4 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 5.8 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 5.8 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 6.3 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 6.3 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 6.7 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 6.7 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 7.2 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 7.2 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 7.6 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 7.7 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 8.0 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 8.1 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 8.5 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 8.6 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 9.0 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 9.0 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 9.4 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 9.4 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 9.5 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 9.9 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 9.9 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 9.9 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 10.3 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 10.3 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 10.4 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 10.8 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 10.8 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 10.8 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 10.8 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 11.2 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 11.2 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 11.2 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 11.7 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 11.7 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 11.7 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 12.1 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 12.1 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 12.1 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 12.6 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 12.6 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 13.0 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 13.1 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 13.1 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 13.5 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 13.5 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 13.5 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 13.9 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 14.0 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 14.0 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 14.4 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 14.4 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 14.4 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 14.4 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 14.8 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 14.8 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 14.8 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 14.8 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 15.3 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 15.3 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 15.3 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 15.3 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 15.7 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 15.7 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 15.7 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 15.7 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 16.2 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 16.2 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 16.2 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 16.2 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 16.6 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 16.6 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 16.7 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 17.1 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 17.1 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 17.1 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 17.5 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 17.5 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 17.6 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 18.0 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 18.0 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 18.4 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 18.4 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 18.5 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 18.9 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 18.9 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 18.9 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 19.3 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 19.3 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 19.4 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 19.8 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 19.8 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 19.8 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 19.8 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 20.2 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 20.2 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 20.2 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 20.7 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 20.7 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 20.7 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 21.1 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 21.1 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 21.1 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 21.6 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 21.6 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 22.0 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 22.1 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 22.5 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 22.5 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 22.9 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 23.0 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 23.4 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 23.4 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 23.8 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 23.8 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 24.3 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 24.3 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 24.7 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 24.7 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 25.2 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 25.2 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 25.6 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 26.1 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 26.5 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000

Conclusions: as can be seen from the data presented in Table 3.1 , the greatest antiviral activity against parainfluenza virus type 3 in vitro on HEp-2 cell culture is exhibited by Trecrezan of compositions (1) and (2) containing the maximum amount of additives, No. 1 - o-cresol , No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc, Moreover, the introduction of any of the tested additives or their set into the Trekrezan preparation, both in the case of composition (1) and in the case of composition (2), leads to an increase in antiviral activity, which is expressed in a decrease in the determined 50% effective concentration.

Table 3.2. − Average results of assessing the antiviral activity according to the chemotherapeutic index (CHI) of the drug Trekrezan (compositions (1) and (2) with additives) against parainfluenza virus type 3 in vitro on HEp-2 cell culture. HTI = CTD ₅₀ /ED ₅₀ Serial numbers of introduced additives No. 1 No. 2 No. 3 No. 4 No. 5 No. 6 No. 7 The concentration of the additive is indicated as a percentage of the active substance contained in the tablet The concentration of the additive is indicated in (mg) per tablet weight 279.3 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 256.0 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 230.2 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 214.6 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 213.7 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 200.2 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 199.3 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 184.8 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 184.1 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 174.3 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 173.6 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 162.5 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 161.8 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 154.3 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 148.4 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 146.4 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 142.1 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 138.7 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 133.7 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 128.7 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 128.0 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 123.4 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 122.9 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 119.6 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 117.4 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 117.1 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 115.6 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 113.2 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 112.7 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 109.9 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 108.2 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 107.7 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 106.6 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 106.0 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 104.6 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 102.9 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 102.5 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 100.0 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 98.7 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 98.5 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 97.4 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 95.8 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 95.4 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 92.2 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 91.7 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 89.5 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 87.1 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 86.6 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 86.1 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 85.0 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 84.7 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 84.2 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 81.9 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 81.6 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 80.2 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 79.8 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 79.8 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 79.4 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 78.2 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 77.8 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 77.8 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 77.4 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 75.8 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 75.6 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 75.4 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 75.2 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 74.1 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 73.8 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 73.8 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 73.4 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 72.0 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 71.7 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 71.6 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 71.3 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 70.4 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 70.1 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 68.2 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 68.2 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 68.0 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 67.1 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 67.1 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 66.8 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 65.1 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 64.1 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 63.8 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 62.8 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 62.5 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 61.3 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 61.2 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 61.1 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 60.3 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 60.2 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 59.9 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 58.8 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 58.7 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 58.5 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 57.9 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 57.8 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 57.6 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 56.9 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 56.6 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 56.3 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 55.6 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 55.6 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 55.4 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 54.7 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 54.5 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 53.6 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 53.3 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 52.7 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 51.5 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 51.2 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 50.9 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 50.6 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 49.4 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 48.9 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 48.7 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 48.2 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 48.0 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 47.3 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 47.2 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 46.7 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 46.5 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 45.9 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 45.7 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 45.2 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 44.3 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 44.0 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000

Conclusions: as can be seen from the data presented in table 3.2 , all obtained CTI values significantly exceed the threshold value of 8, which indicates the presence of antiviral activity of Trekrezan compositions (1) and (2) with any combination of additives, while the greatest antiviral activity against parainfluenza virus type 3 in vitro on HEp-2 cell culture exhibits Trecrezan of both compositions (1) and (2) containing the maximum amount of additives, No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-( 3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc , while introducing any of the tested additives or a set of them into the Trekrezan preparation as in the case composition (1), and in the case of composition (2) leads to an increase in antiviral activity, which is expressed in an increase in the estimated CTI index .

The results of the study of the antiviral activity of the drug Trecrezan composition (1) and (2) without additives against coronavirus OS43 are presented in average form in Table 4.

Table 4 – Results of testing the antiviral activity of the drug Trekrezan against coronavirus OS43 in vitro Concentration, µg/ml Virus titer, lg TID ₅₀ Decrease in titer compared to control Decrease in infectious activity compared to control, % ED ₅₀ 300 2.17±0.76 1.50 96.84 90.2 100 3.17±0.76 0.50 68.38 33 3.17±0.76 0.50 68.38 eleven 3.5±0.5 0 0 3.3 3.67±0.29 - - 0 3.5±0.5 - -

From the data presented in the table it can be seen that the ED value of ₅₀ Trecrezan preparations of composition (1) and (2) without additives for coronavirus OS43 was 12.2. This results in a value of HTI = 1374/90.2 = 15.2, which exceeds the threshold value of 8.

Additionally, experiments were carried out with the drug Trekrezan of compositions (1) and (2) with the introduction of different concentrations and sets of additives, No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid , No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc. The results of determining the antiviral activity according to ED ₅₀ (50% effective dose) of the drug Trekrezan (compositions (1) and (2) with additives) are presented in average form in Tables 4.1-4.2.

Table 4.1 - Average results of determining the antiviral activity according to ED ₅₀ (50% effective dose) of the drug Trecrezan (compositions (1) and (2) with additives) against coronavirus type OC43 in vitro on Vero cell culture. ED ₅₀ , mcg/ml Serial numbers of added additives No. 1 No. 2 No. 3 No. 4 No. 5 No. 6 No. 7 The concentration of the additive is indicated as a percentage of the active substance contained in the tablet The concentration of the additive is indicated in (mg) per tablet weight 13.8 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 15.4 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 16.9 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 18.4 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 18.4 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 19.9 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 19.9 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 21.4 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 21.4 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 22.9 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 22.9 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 24.4 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 24.4 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 25.9 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 26.1 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 27.4 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 27.6 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 28.9 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 29.1 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 30.6 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 30.6 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 32.1 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 32.1 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 32.2 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 33.6 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 33.6 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 33.7 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 35.1 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 35.1 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 35.2 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 36.6 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 36.6 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 36.8 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 36.8 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 38.2 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 38.3 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 38.3 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 39.7 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 39.8 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 39.8 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 41.2 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 41.3 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 41.3 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 42.8 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 42.8 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 44.3 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 44.5 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 44.5 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 45.8 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 46.0 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 46.0 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 47.3 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 47.5 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 47.5 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 49.0 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 49.0 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 49.0 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 49.0 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 50.5 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 50.5 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 50.5 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 50.5 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 52.0 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 52.0 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 52.0 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 52.0 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 53.5 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 53.5 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 53.5 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 53.5 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 55.0 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 55.0 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 55.0 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 55.0 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 56.5 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 56.5 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 56.8 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 58.0 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 58.0 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 58.3 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 59.6 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 59.6 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 59.8 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 61.3 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 61.3 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 62.8 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 62.8 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 62.9 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 64.3 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 64.3 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 64.4 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 65.8 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 65.8 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 65.9 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 67.3 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 67.3 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 67.4 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 67.4 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 68.8 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 68.9 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 68.9 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 70.3 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 70.4 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 70.4 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 71.8 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 71.9 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 71.9 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 73.4 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 73.4 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 74.9 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 75.1 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 76.4 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 76.6 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 77.9 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 78.2 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 79.7 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 79.7 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 81.2 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 81.2 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 82.7 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 82.7 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 84.2 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 84.2 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 85.7 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 85.7 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 87.2 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 88.7 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 90.2 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000

Conclusions: as can be seen from the data presented in Table 4.1 , the greatest antiviral activity against type OC43 coronavirus in vitro on Vero cell culture is exhibited by Trecrezan of compositions (1) and (2) containing the maximum amount of additives, No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc, with the introduction any of the tested additives or their set in the Trekrezan drug, both in the case of composition (1) and in the case of composition (2), leads to an increase in antiviral activity, which is expressed in a decrease in the determined 50% effective concentration.

Table 4.2. − Average results of assessing the antiviral activity according to the chemotherapeutic index (CHI) of the drug Trekrezan (compositions (1) and (2) with additives) against coronavirus OS43 in vitro on Vero cell culture. HTI = CTD ₅₀ /ED ₅₀ Serial numbers of added additives No. 1 No. 2 No. 3 No. 4 No. 5 No. 6 No. 7 The concentration of the additive is indicated as a percentage of the active substance contained in the tablet The concentration of the additive is indicated in (mg)
per tablet weight 98.0 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 88.4 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 80.5 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 74.5 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 74.1 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 69.0 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 68.6 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000050 64.3 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000050 64.1 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 60.3 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 60.0 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000050 56.7 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000050 56.4 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000050 53.5 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000050 51.8 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 50.5 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000050 49.3 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 48.2 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000050 46.7 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 44.7 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 44.5 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 42.7 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 42.5 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000050 41.6 0.5000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 40.9 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000050 40.7 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 40.1 0.5000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 39.3 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 39.1 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000050 38.3 0.5000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 37.7 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000050 37.6 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000050 36.9 0.5000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 36.7 0.5000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 36.2 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000050 35.5 0.5000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 35.4 0.0000 0.5000 0.5000 0.5000 0.7500 0.29000 0.000000 34.8 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000050 34.3 0.0000 0.0000 0.5000 0.5000 0.7500 0.29000 0.000000 34.2 0.5000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 33.8 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000050 33.1 0.5000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 33.0 0.0000 0.5000 0.5000 0.0000 0.7500 0.29000 0.000000 32.0 0.0000 0.0000 0.5000 0.0000 0.7500 0.29000 0.000000 31.9 0.0000 0.5000 0.0000 0.5000 0.7500 0.29000 0.000000 31.0 0.0000 0.0000 0.0000 0.5000 0.7500 0.29000 0.000000 30.3 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 30.1 0.5000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 30.0 0.0000 0.5000 0.0000 0.0000 0.7500 0.29000 0.000000 29.5 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 29.3 0.5000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 29.2 0.0000 0.0000 0.0000 0.0000 0.7500 0.29000 0.000000 28.5 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 28.4 0.5000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 27.8 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 27.7 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 27.7 0.5000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 27.6 0.5000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 27.1 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 27.0 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000050 26.9 0.5000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 26.8 0.0000 0.5000 0.5000 0.5000 0.0000 0.29000 0.000000 26.3 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 26.3 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000050 26.2 0.0000 0.0000 0.5000 0.5000 0.0000 0.29000 0.000000 26.1 0.5000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 25.7 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 25.6 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000050 25.6 0.5000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 25.4 0.0000 0.5000 0.5000 0.0000 0.0000 0.29000 0.000000 25.0 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000050 24.9 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000050 24.9 0.0000 0.0000 0.5000 0.0000 0.0000 0.29000 0.000000 24.8 0.0000 0.5000 0.0000 0.5000 0.0000 0.29000 0.000000 24.4 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000050 24.3 0.0000 0.0000 0.0000 0.5000 0.0000 0.29000 0.000000 23.8 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000050 23.7 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 23.6 0.0000 0.5000 0.0000 0.0000 0.0000 0.29000 0.000000 23.3 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000050 23.2 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 23.2 0.0000 0.0000 0.0000 0.0000 0.0000 0.29000 0.000000 22.6 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 22.2 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 22.1 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 21.7 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 21.6 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000050 21.3 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000050 21.2 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 21.2 0.5000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 20.9 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 20.9 0.5000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 20.8 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000050 20.4 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000050 20.4 0.5000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 20.3 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000050 20.1 0.5000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 20.0 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000050 20.0 0.5000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 19.7 0.5000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 19.6 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000050 19.6 0.0000 0.5000 0.5000 0.5000 0.7500 0.00000 0.000000 19.3 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000050 19.3 0.0000 0.0000 0.5000 0.5000 0.7500 0.00000 0.000000 19.2 0.5000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 19.0 0.5000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 18.9 0.0000 0.5000 0.5000 0.0000 0.7500 0.00000 0.000000 18.6 0.0000 0.0000 0.5000 0.0000 0.7500 0.00000 0.000000 18.5 0.0000 0.5000 0.0000 0.5000 0.7500 0.00000 0.000000 18.3 0.0000 0.0000 0.0000 0.5000 0.7500 0.00000 0.000000 17.9 0.0000 0.5000 0.0000 0.0000 0.7500 0.00000 0.000000 17.8 0.5000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 17.7 0.0000 0.0000 0.0000 0.0000 0.7500 0.00000 0.000000 17.5 0.5000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 17.1 0.5000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 16.9 0.5000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 16.9 0.5000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 16.7 0.5000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 16.6 0.0000 0.5000 0.5000 0.5000 0.0000 0.00000 0.000000 16.4 0.0000 0.0000 0.5000 0.5000 0.0000 0.00000 0.000000 16.3 0.5000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 16.2 0.5000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000 16.1 0.0000 0.5000 0.5000 0.0000 0.0000 0.00000 0.000000 15.9 0.0000 0.0000 0.5000 0.0000 0.0000 0.00000 0.000000 15.8 0.0000 0.5000 0.0000 0.5000 0.0000 0.00000 0.000000 15.7 0.0000 0.0000 0.0000 0.5000 0.0000 0.00000 0.000000 15.4 0.0000 0.5000 0.0000 0.0000 0.0000 0.00000 0.000000 15.2 0.0000 0.0000 0.0000 0.0000 0.0000 0.00000 0.000000

Conclusions: as can be seen from the data presented in Table 4.2 , all obtained CTI values significantly exceed the threshold value of 8, which indicates the presence of antiviral activity of Trekrezan compositions (1) and (2) with any combination of additives, while the greatest antiviral activity against coronavirus OS43 in vitro on Vero cell culture exhibits Trecrezan of both compositions (1) and (2), containing the maximum amount of additives, No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy ) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc, while introducing any of the tested additives or a set of them into the Trekrezan preparation as in the case of composition ( 1), and in the case of composition (2) leads to an increase in antiviral activity, which is expressed in an increase in the estimated CTI index.

Another unexpected technical result turned out to be that the claimed pharmaceutical composition for obtaining a medicinal product in solid dosage form based on hydroxyethylammonium methylphenoxyacetate with additional components provides high bioavailability due to faster entry of the active component into the blood and, accordingly, ensures a faster onset of the therapeutic effect, due to which is more promising for obtaining a drug with immunostimulating and antiviral properties.

Brief description of drawings

Figure 1 shows six curves - averaged dissolution profiles of the drug Trecrezan compositions (1) and (2), containing different combinations of additives, namely, curve No. 1 (without additives *), curve No. 2 (with additives No. 1 and 5 *) , curve No. 3 (with additives No. 5, 6 and 7*), curve No. 4 (with additives No. 1, 5, 6 and 7*), curve No. 5 (with additives No. 1, 2, 5, 6 and 7* ), curve No. 6 (with all seven additives*) (medium - 0.1 M hydrochloric acid solution).

*list of additives No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc.

In FIG. Figure 2 shows six curves - averaged dissolution profiles of the drug Trekrezan compositions (1) and (2), containing different combinations of additives, namely, curve No. 1 (without additives *), curve No. 2 (with additives No. 1 and 5 *), curve No. 3 (with additives No. 5, 6 and 7*), curve No. 4 (with additives No. 1, 5, 6 and 7*), curve No. 5 (with additives No. 1, 2, 5, 6 and 7*), curve No. 6 (with all seven additives*) (medium - acetate buffer solution pH 4.5).

*list of additives No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc.

In FIG. Figure 3 shows six curves - averaged dissolution profiles of the drug Trekrezan compositions (1) and (2), containing different combinations of additives, namely, curve No. 1 (without additives *), curve No. 2 (with additives No. 1 and 5 *), curve No. 3 (with additives No. 5, 6 and 7*), curve No. 4 (with additives No. 1, 5, 6 and 7*), curve No. 5 (with additives No. 1, 2, 5, 6 and 7*), curve No. 6 (with all seven additives*) (phosphate buffer solution pH 6.8).

*list of additives No. 1 - o-cresol, No. 2 - 2-phenoxyacetic acid, No. 3 - 2-(3-methylphenoxy) acetic acid, No. 4 - 2-(4-methylphenoxy) acetic acid, No. 5 - calcium, No. 6 - phosphorus and No. 7 - zinc.

In FIG. 4 shows the scheme of the clinical trial

Carrying out the invention

The proposed solid pharmaceutical composition is prepared as follows.

Example 1

If necessary, grind the required amount of the active components of hydroxyethylammonium methylphenoxyacetate, o-cresol, 2-phenoxyacetic acid, 2-(3-methylphenoxy) acetic acid, 2-(4-methylphenoxy) acetic acid or a mixture of these substances in a mill with a mesh size of 0. 8-1.5 mm.

The ground substances are collected in production containers and sent to the sieving operation.

Sift the required amount of ingredients through a sieve with a size of: 0.5-1.0 mm – crushed hydroxyethylammonium methylphenoxyacetate, microcrystalline cellulose; lactose monohydrate, potato starch, calcium stearate, zinc/calcium chelate, phosphoric acid salt, sodium stearyl fumarate; 0.8-1.0 mm – colloidal silicon dioxide (aerosil).

After sifting, the powders are collected in production containers and sent to the stage of obtaining the mass for the dosage form by direct compression or wet granulation, from which the drug is obtained in tablet form. If the drug is obtained in capsule form, the sifted powders are filled into capsules.

The study of dissolution kinetics was carried out in accordance with GPM.1.4.2.0014.15 “Dissolution for solid dosage forms” on a “rotating basket” apparatus (Erweka DT 126 Light, Germany) at a rotation speed of 100 rpm.

Example 2

The proposed solid pharmaceutical composition is administered orally:

- for the prevention of acute respiratory viral infections as part of complex therapy: adults and children over 12 years of age are prescribed 1 tablet (200 mg oxyethylammonium methylphenoxyacetate) per day after meals. A total of 14 tablets per course (total dose 2800 mg hydroxyethylammonium methylphenoxyacetate), course duration 14 days;

- for the treatment of acute respiratory viral infections and influenza as part of complex therapy: adults and children over 12 years of age are prescribed on the first day 1 tablet 3 times (600 mg oxyethylammonium methylphenoxyacetate) per day, in the next 7 days 1 tablet (200 mg hydroxyethylammonium methylphenoxyacetate ). A total of 10 tablets per course (total dose 2000 mg), course duration 8 days;

- during periods of high intellectual and heavy physical activity, as part of complex therapy, adults and children over 12 years of age are prescribed 3 tablets (600 mg) per day on the first day, 1 tablet (200 mg hydroxyethylammonium methylphenoxyacetate) for the next 7 days. A total of 10 tablets per course (total dose 2000 mg), course duration 8 days;

- to increase the body’s resistance to various stressful influences (hypoxia, overheating, hypothermia) and adverse environmental influences (sharp changes in climatic conditions, adaptation to changes in atmospheric pressure), as part of complex therapy for adults and children over 12 years of age, 3 tablets are prescribed on the first day ( 600 mg) per day, in the next 7 days, 1 tablet (200 mg). A total of 10 tablets per course (total dose 2000 mg hydroxyethylammonium methylphenoxyacetate), course duration 8 days.

Example 3

To confirm the presence of adaptogenic, immunomodulatory and antiviral effects in patients, a double-blind, randomized, placebo-controlled comparative multicenter parallel study of the effectiveness and safety of the proposed composition in the treatment of influenza was conducted.

A particular version of the proposed composition (wt.%) was chosen for the study:

hydroxyethylammonium methylphenoxyacetate 40 o-cresol 0.1 2-(3-methylphenoxy)acetic acid 0.002 Calcium 0.25 Zinc 0.00034 Excipients up to 100.

Microcrystalline cellulose, lactose monohydrate, sodium stearyl fumarate, colloidal silicon dioxide (Aerosil), and potato starch were used as excipients.

The purpose of the study was to evaluate the effectiveness of the proposed composition in comparison with the drug Trekrezan®, tablets, 200 mg (Grotex LLC, Russia) in the treatment of influenza, to evaluate the safety and tolerability of the drug in the treatment of influenza.

Research objectives:

To evaluate the effectiveness of the proposed composition in comparison with the drug Trekrezan®, tablets, 200 mg (Grotex LLC, Russia) in the treatment of influenza (according to examination and patient diary), based on:

the proportion of patients with normalization of morning body temperature on the 5th day of therapy;

proportions of patients with improvement on days 3, 5, 7, 9 of therapy;

proportions of patients with recovery on days 3, 5, 7, 9 of therapy;

severity of influenza (total influenza severity index) on days 3, 5, 7, 9 of therapy;

severity of clinical manifestations of influenza (fever, general symptoms and symptoms of the nose/throat/chest) in points) on days 3, 5, 7, 9 of therapy;

duration of the main clinical symptoms of influenza (fever, general symptoms and symptoms of the nose/throat/chest) in days;

the proportion of patients who required the use of antipyretic drugs on days 3, 5, 7, 9 of therapy;

the proportion of patients who developed complications requiring antibiotics;

the proportion of patients with a negative virological test result on day 5 of therapy (based on PCR results of samples from the nose and oropharynx);

changes in immune status indicators on day 5 of therapy.

To evaluate the safety and tolerability profile of the proposed composition and the drug Trekrezan®, tablets, 200 mg (Grotex LLC, Russia) (incidence of adverse events (AEs) and serious adverse events (SAEs), changes in laboratory test data, vital parameters ( ZhVP)).

The criterion for absence of fever is temperature below 37.0°C for 24 hours

Improvement criteria: body temperature ≤37.2°C + total severity of all symptoms (except fever index) ≤2 points.

Recovery criteria: body temperature < 37.0°C + absence of symptoms

To assess temperature in points for the purpose of calculating the total disease index, absolute values of body temperature measured in degrees Celsius will be converted into relative units (or points) taking into account the following gradations: ≤37.2 ° C = 0 points; 37.3-38.0°C=1 point; 38.1-39.0°C=2 points; ≥39.1°C=3 points.

One hundred and forty (140) adult patients with influenza confirmed by a positive rapid influenza diagnostic test, male and female, aged 18 to 65 years (inclusive), selected according to inclusion criteria and without exclusion criteria, were included in the study.

Duration of the study: the total duration of the study for each patient was no more than 13 days:

Screening stage:

Visit 0, day 1 – outpatient visit;

Therapy stage:

Visit 1, day 1 – outpatient visit, randomization and initiation of therapy;

Visit 2, day 3 – telephone contact (TC) for the purpose of safety assessment;

Visit 3, day 5 – outpatient visit;

Visit 4, day 7±1 – TC to assess safety;

Visit 5, day 9 – outpatient visit, completion of therapy;

Observation stage:

Visit 6, day 12±1 – TC to assess safety.

Due to the fact that the study included patients with a fever of 37.8°C or higher, as well as severe (≥4 points on the symptom rating scale) clinical manifestations of influenza, home visits were allowed.

GENERAL PLAN (DESIGN) OF THE STUDY: Basic and additional parameters under study

Performance evaluation criteria

Primary endpoint:

• the proportion of patients with normalization of morning body temperature on the 5th day of therapy;

Secondary endpoints:

• proportion of patients with improvement on days 3, 5, 7, 9 of therapy;

• the proportion of patients with recovery on days 3, 5, 7, 9 of therapy;

• severity of influenza (total influenza severity index) on days 3, 5, 7, 9 of therapy;

• severity of clinical manifestations of influenza (fever, general symptoms and symptoms of the nose/throat/chest) in points) on days 3, 5, 7, 9 of therapy;

• duration of the main clinical symptoms of influenza (fever, general symptoms and symptoms of the nose/throat/chest) in days;

• the proportion of patients who required the use of antipyretic drugs on days 3, 5, 7, 9 of therapy;

• the proportion of patients who developed complications requiring antibiotics;

• the proportion of patients with a negative virological test result on day 5 of therapy (based on PCR results of samples from the nose and oropharynx)

• dynamics of immune status indicators on the 5th day of therapy.

Safety and tolerability indicators

The safety of the compared drugs will be assessed according to the following parameters:

1. Assessment of AEs and SAEs: all information on AEs/SAEs, as well as information on concomitant therapy for the treatment of AEs/SAEs.

2. Changes to inspection and research data:

• changes in indicators based on the results of assessing basic vital parameters (SBP, DBP, RR, heart rate);

• changes in laboratory examination data (UAC, OAM, BhAC);

Tolerability will be assessed by the proportion of patients who discontinued the study early due to AEs/SAEs.

Description of the type (design) of the study being conducted

Type of study

A phase IV study is planned. In accordance with International Conference on Harmonization E8 guidelines, the purpose of this study is to provide additional data regarding the drug's efficacy, safety and tolerability compared with placebo in the treatment of influenza (new indication).

A graphical diagram of the clinical study is presented in Figure 4.

The effectiveness of therapy is assessed based on:

TT (in degrees Celsius);

general symptoms – headache, chills, sweating, weakness, muscle pain, drowsiness;

Nasal symptoms – nasal discharge, nasal congestion, sneezing;

throat symptoms - sore throat, hoarseness/hoarseness of the voice;

chest symptoms - cough, chest pain/heaviness;

• a test for determining the antigens of influenza viruses type A and B using a qualitative immunochromatographic rapid test;

• laboratory research:

PCR diagnostics of smears from the mucous membrane of the nose and oropharynx (nasopharyngeal smears) (detection of RNA viruses of influenza A (Influenza virus A) and influenza B (Influenza virus B));

general urine analysis with microscopic examination (OAM) (color, transparency, urine pH, specific gravity, nitrites, protein, glucose, ketones, urobilinogen, bilirubin, microscopic examination (erythrocytes, leukocytes, casts, except hyaline, bacteria));

complete blood count (CBC) (hemoglobin, red blood cells, hematocrit, platelets, leukocytes (abs., relative), erythrocyte sedimentation rate (ESR), leukocyte count (abs., relative) (neutrophils, lymphocytes, eosinophils, monocytes, basophils));

biochemical blood test (BhAK) (total protein, creatinine, urea, glucose, total bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total cholesterol);

immunological blood test (IAB) (serum interferon (IFN)), level of spontaneous and induced production of IFN-α and IFN-γ by leukocytes);

List of efficiency parameters

Primary endpoint:

• the proportion of patients with normalization of morning body temperature on the 5th day of therapy;

Secondary endpoints:

• proportion of patients with improvement on days 3, 5, 7, 9 of therapy;

• the proportion of patients with recovery on days 3, 5, 7, 9 of therapy;

• severity of influenza (total influenza severity index) on days 3, 5, 7, 9 of therapy;

• severity of clinical manifestations of influenza (fever, general symptoms and symptoms of the nose/throat/chest) in points) on days 3, 5, 7, 9 of therapy;

• duration of the main clinical symptoms of influenza (fever, general symptoms and symptoms of the nose/throat/chest) in days;

• the proportion of patients who required the use of antipyretic drugs on days 3, 5, 7, 9 of therapy;

• the proportion of patients who developed complications requiring antibiotics;

• the proportion of patients with a negative virological test result on day 5 of therapy (based on PCR results of samples from the nose and oropharynx);

• dynamics of immune status indicators on the 5th day of therapy.

Recovery criteria: body temperature <37.0°C + absence of symptoms.

Improvement criteria: body temperature ≤37.2°C + total severity of all symptoms (except fever index) ≤2 points.

Fever-free criterion: temperature below 37.0°C for 24 hours.

According to the recommendations (Ministry of Health of the Russian Federation, 2014), it is recommended to use rapid methods for detecting viral antigens (including immunochromatographic) as simple and convenient methods for mass immunodiagnosis, accessible to widespread practice.

Dosage regimen for study drugs:

Duration of therapy 8 days

Therapy groups:

• Group I (G1, PI) – study group, whose patients will receive the proposed composition according to the treatment regimen;

• Group II (G2, PS) – control group, patients of which will receive Trekrezan®, tablets, 200 mg (Grotex LLC, Russia) according to the treatment regimen.

Treatment regimen:

• on the first day, 1 tablet 3 times a day;

• in the next 7 days, 1 tablet 1 time/day;

• a total of 10 tablets per course;

• course duration is 8 days.

Throughout the study, patients can receive symptomatic treatment for influenza, based on accepted standards of treatment, including detoxification agents, vitamins, antitussives or expectorants, mucolytics, vasoconstrictor nasal drops, and, if indicated, antipyretic drugs.

Indications for prescribing antipyretic drugs: increased body temperature above 39°C in patients without complications or concomitant diseases

Drugs approved for use as antipyretics (ATC group): Paracetamol (N02BE01), Ibuprofen (M01AE01). The choice of drug is made by the study physician, who gives the antipyretic drug to the patient at Visit 1.

Blinding, randomization:

Blinding of comparison groups is provided: the study proposed composition and the drug Trekrezan®, tablets, 200 mg (Grotex LLC, Russia).

Thus, neither the doctor nor the patient will know which drug is prescribed.

Criteria for assessing safety and tolerability:

The safety profile is assessed by the frequency of occurrence of AEs/SAEs, changes in laboratory data (CBC, BHA, OAM), changes in these functions of vital organs and gastrointestinal tract (HR, RR, TT, SBP, DBP).

Based on the results of clinical studies, the effectiveness of the proposed composition was proven in comparison with the drug Trekrezan without additives in the treatment of influenza; an improvement in the condition was already observed on the 3rd day compared with the drug Trekrezan without additives, and complete recovery on the 9th day, while when taking With the drug Trecrezan without additives, residual effects of influenza were observed on the 9th day in the form of residual runny nose and cough. It has also been proven that the proposed composition helps to increase the spontaneous and induced production of alpha and gamma interferons, which is manifested in its adaptogenic and immunomodulatory properties.

Based on the studies conducted, it is clear that the new adaptogenic, immunomodulatory and antiviral pharmaceutical composition for obtaining a solid drug based on oxyethylammonium methylphenoxyacetate with additional components has faster dissolution profiles and thereby ensures a faster entry of medicinal substances into the blood and, accordingly, ensures a faster onset of therapeutic effect.

Claims (9)

1. Solid pharmaceutical composition with adaptogenic, immunomodulatory and antiviral properties against influenza A, influenza B, parainfluenza and seasonal coronavirus OS43 viruses, containing oxyethylammonium methylphenoxyacetate, additional components and excipients in the following ratio of components, wt. %: hydroxyethylammonium methylphenoxyacetate 20-60 o-cresol 0.00001-0.5 and/or 2-phenoxyacetic acid and/or 2-(3-methylphenoxy) acetic acid and/or 2-(4-methylphenoxy) acetic acid or a mixture of these substances 0.00001-0.5 Phosphorus 0.00001-0.29 and/or Zinc 0.0000001-0.00050 or a mixture thereof microcrystalline cellulose, lactose monohydrate, sodium stearyl fumarate, calcium stearate, aerosil, potato starch or mixtures thereof up to 100. 2. Solid pharmaceutical composition according to claim 1, characterized in that the composition is obtained from the substance hydroxyethylammonium methylphenoxyacetate, which has a crystalline structure. 3. Solid pharmaceutical composition according to claim 1, characterized in that it is made in the dosage form of tablets or capsules. 4. Solid pharmaceutical composition according to claim 1, characterized in that it can be used in conjunction with etiotropic therapy drugs.