RU2800287C1 - Antibacterial composition of gramicidin c, a method of its production and use - Google Patents

Abstract

FIELD: medicine and chemical-pharmaceutical industry.

SUBSTANCE: group of inventions concerns new medicines based on gramicidin C. The antibacterial complex consists of 2-hydroxypropyl-β-cyclodextrin, gramicidin C or its pharmaceutically acceptable salt (in terms of gramicidin C), cetylpyridinium chloride or its monohydrate (in terms of cetylpyridinium chloride) in a mass ratio of 6-100:3:1. The antibacterial composition for preparing a solution for local or external use is made in the form of a tablet and contains the indicated antibacterial complex as an active principle and mannitol, crospovidone, sorbitol, aspartame and polyethylene glycol 4,000 as auxiliary components. The components are used in the declared quantities. In another embodiment, a method of preparing the said composition is provided. The specified composition is used to obtain an antibacterial agent of local or external action in the form of an aqueous solution.

EFFECT: use of the group of inventions ensures the production of a new stable antibacterial composition based on the gramicidin C complex, which has improved solubility, as well as helps to increase the effectiveness of the antibacterial effect while minimizing the toxic effect on the human body and microbiota.

6 cl, 3 tbl, 4 ex

Description

The invention relates to the field of medicine and pharmaceutical industry and relates to new drugs based on gramicidin C.

Prior Art

One of the popular and effective therapy for patients with infectious and inflammatory diseases of the oral cavity is the use of topical drugs, which minimizes the toxic effect on the human body and microbiota (Karneeva O.V., Daihes N.A., Polyakov D.P. Protocols for the diagnosis and treatment of acute tonsillopharyngitis // Russian Medical Journal. 2015. No. 6. P. 307-311).

Patients with infectious and inflammatory diseases of the pharynx are recommended to prescribe ^Grammidin® neo or Grammidin® with neo anesthetic (Svistushkin V.M., Nikiforova G.N., Mokoyan Zh.T. The place of topical antimicrobial drugs in the pharmacotherapy of infectious and inflammatory diseases of the pharynx. RMJ. 2017; 23:1739-1743). The active substances of the drug Grammidin ^® neo are gramicidin C and cetylpyridinium chloride.

Gramicidin C was first obtained in the USSR in 1942. It is a cyclic decapeptide containing two amino acid residues: L-valine, L-ornithine, L-leucine, D-phenylalanine and L-proline. Gramicidin C has an antibacterial effect against a wide range of gram-positive and gram-negative microorganisms.

Technical solutions are known in which gramicidin C is in solid form, drugs with it are produced in the form of lozenges. (Patents of the Russian Federation No. 2213558, 2160088, 2590980). However, gramicidin C is practically insoluble in water, which reduces its bioavailability, so various methods have been proposed to increase its solubility.

Known pharmaceutical substance based on gramicidin C, cyclodextrin and chitosan, in which gramicidin C is encapsulated in cyclodextrin with subsequent formation of a polyelectrolyte complex with chitosan (RF Patent No. 2733715). The components are in the following ratios in wt %: gramicidin C (1-31.25), cyclodextrin (2-62.5) and chitosan (6.25-97). The pharmaceutical substance has a prolonged action and increased solubility due to the possible encapsulation of the most hydrophobic part of D-phenylalanine in the gramicidin C molecule. At the same time, the technical effect of this technical solution is to expand the means of this purpose, and exact data on the increase in solubility are not given.

In another work, it was shown that when comparing the chemical shifts of gramicidin C, cyclodextrin and their joint complex, a significant change in the chemical value from 7.91 to 7.69 was found for the protons of the amino group of L-ornithine in the gramicidin C molecule in a complex with cyclodextrin (Drannikov A. A., Vatlin I. S., Trusova M. E., Di Martino A., Krivoshchekov S. V., Guriev A. M., Belousov M. V. Investigation of the effect of the method of obtaining inclusion complexes of gramicidin C and β-cyclodextrin on their technological performance. Development and registration of drugs. 2022; 11(2) pp. 102-108. https://doi.org/10.33380/2305-2066-2022-11-2-102-108).

Cyclodextrins are derivatives of starch and are widely used in the pharmaceutical industry due to their specific structure with a cavity suitable for incorporating other molecules. This specific property of cyclodextrins is widely used in pharmacy to change the release of the drug, mask the negative organoleptic properties, as well as β to improve its solubility in water (RF Patent No. 2099354). However cyclodextrins, both natural, such as α, β, γ, and various synthetic derivatives, such as hydroxypropyl derivatives of β, γ and methylated β-cyclodextrins, sulfobutyl ethers of β-cyclodextrin, etc., differing in structure, respectively, in properties and application, do not provide an equivalent improvement in the necessary properties for different pharmaceutically active substances.

In particular, complexation of gramicidin C with β-cyclodextrin was found to increase the aqueous solubility of the drug and provide a sustained release of gramicidin C at pH = 4.3 for at least 48 h. chemical technology in the XXI century: materials of the XXI International scientific and practical conference of students and young scientists named after outstanding chemists L. P. Kulev and N. M. Kizhner, dedicated to the 110th anniversary of the birth of Professor A. G. Stromberg, September 21-24, 2020, Tomsk Polytechnic University, pp. 525-526.).

The disadvantage of the complex of gramicidin C with β-cyclodextrin is the lack of release of gramicidin C at pH above 4.3, which makes it unsuitable for almost all physiological media, including when it is dissolved in ordinary water with a neutral pH for gargling.

Liquid dosage forms based on gramicidin C and cetylpyridinium chloride are known (RF patents Nos. 2627423, 2749902). These compositions include ethanol in their composition, which significantly limits their use by drivers of vehicles, pregnant women and other categories of citizens.

The closest in composition to the claimed antibacterial composition is the drug Grammidin ^® neo in the form of lozenges with antibacterial action, the composition of which is taken as a prototype (https://www.vidal.ru/drugs/grammidin_neo_20563).

Grammidin ^® neo has the following composition of substances in the composition:

gramicidin C dihydrochloride - 3.20 mg (in terms of gramicidin C - 3 mg);

cetylpyridinium chloride monohydrate - 1.05 mg (in terms of cetylpyridinium chloride - 1 mg);

colloidal silicon dioxide - 5 mg;

talc - 60 mg;

acesulfame potassium - 15 mg;

mint flavor - 25 mg;

sorbitol - 1065.75 mg;

magnesium stearate - 25 mg.

The total weight of the tablet is 1200 mg.

This drug has found wide application in medical practice for the treatment of infectious and inflammatory diseases of the oral cavity and pharynx, tonsillitis, pharyngitis, tonsillitis, periodontal disease, gingivitis and stomatitis. Grammidin ^® neo is a combined drug that reduces inflammation, has an antimicrobial effect, softens discomfort in the throat, facilitates swallowing, causes hypersalivation during resorption, which contributes to the mechanical cleansing of the oral cavity and pharynx from microorganisms.

The present invention, in contrast to the prototype, is a tablet for preparing a solution for topical and external use. Replacement of lozenges with mouthwash is considered a medically and economically effective preventive measure for upper respiratory tract infections (Sakai M., Shimbo T., Omata K. et al.; Great Cold Investigators-I. Cost-effectiveness of gargling for the prevention of upper respiratory tract infections. BMC Health Serv. Res. 2008; 8: P. 258). During irrigation procedures, a significant area of contact of the solution with the mucous membrane arises, favorable conditions are created for a better local effect (E.V. Nosulya. Rinsing in the complex treatment of tonsillopharyngitis. Therapeutic archive 10, 2014, p. 128-131). When rinsing, cleansing occurs more effectively due to respiratory movements with liquid in the oral cavity, the intensity of which is regulated by the patient himself. It also expands the range of application of the solution, which can be used not only for gargling, but for external purposes as a local antibacterial agent.

The difficulty of obtaining such a solution is the almost lack of solubility of gramicidin C in water. The solution to this problem could be to increase the solubility of gramicidin C in water with a pH of 5.5 or more.

The objective of the proposed technical solution is to develop an antibacterial complex of gramicidin C with improved solubility and a new stable antibacterial composition in the form of tablets for preparing a solution for topical or external use based on it.

The problem is solved by the fact that it was possible to achieve increased solubility of gramicidin C in water by including gramicidin C or its pharmaceutically acceptable salt and cetylpyridinium chloride or its monohydrate, which is a surfactant, in the matrix of 2-hydroxypropyl-β-cyclodextrin. This combination of three components in the form of the claimed gramicidin C complex made it possible to increase its solubility to a degree sufficient to prepare an effective antibacterial solution for topical or external use.

Gramicidin C and its pharmaceutically acceptable salts have a pronounced antimicrobial effect against pathogens of infectious diseases of the oral cavity and pharynx. Gramicidin C can be used in the form of pharmaceutically acceptable salts, such as hydrochlorides, hydrobromides, sulfates, phosphates, benzoates, lactates, citrates, maleates, fumarates, succinates, tartrates, malonates, salicylates, etc. (RU 2749902 C2). As the preferred salt, the dihydrochloride salt may be mentioned.

Cetylpyridinium chloride (cetylpyridinium chloride monohydrate) is an antiseptic from the group of quaternary ammonium bases, has variable antifungal activity, bactericidal action against gram-positive and gram-negative bacteria, and is effective against some viruses. Cetylpyridinium chloride has a disinfectant effect on the mucous membrane.

2-hydroxypropyl-β-cyclodextrin is a cyclic oligosaccharide whose molecules are shaped like a torus or a hollow truncated cone. There are no hydroxyl groups inside the torus, so the inner cavity is hydrophobic, and the outer shell is hydrophilic. Surprisingly, the choice of this cyclodextrin in combination with cetylpyridinium chloride made it possible to overcome the problems associated with the dependence of the solubility of gramicidin C on pH.

The antibacterial complex according to the present invention includes 2-hydroxypropyl-β-cyclodextrin, gramicidin C or its pharmaceutically acceptable salt (in terms of gramicidin C), cetylpyridinium chloride or its monohydrate (in terms of cetylpyridinium chloride) in mass ratios of 6-100:3:1. The preferred ratio is 25:3:1.

The antibacterial pharmaceutical composition, made in the form of a tablet for preparing a solution for local or external use, includes the specified complex and a set of pharmaceutically acceptable excipients.

In more detail, the antibacterial pharmaceutical composition, in the form of a tablet for preparing a solution for topical or external use, has the following composition:

antibacterial complex 10.00-104.00 mg

mannitol - 230.0-270.0 mg

crospovidone - 30.0-36.0 mg

sorbitol - 29.0-35.0 mg

aspartame - 6.0-10.0 mg;

polyethylene glycol 4000 -3.0-5.0 mg.

Tablets may additionally include at least one flavor, preferably in an amount of 6-13 mg. For example, at least one flavor may be selected from the group: raspberry, strawberry, apricot, mint, pear, apple, lemon, cinnamon, and the like.

Another object of the invention is a method for obtaining the claimed antibacterial composition, which is carried out in several stages.

At the first stage, the necessary weights of cetylpyridinium chloride (or its monohydrate) and 2-hydroxypropyl-β-cyclodextrin are taken, water is added, stirred until a clear solution is formed, gramicidin C or its pharmaceutically acceptable salt is added to it, stirred for 40-50 minutes until a transparent or slightly opalescent solution is obtained, the resulting solution is frozen and lyophilized. A complex is formed from gramicidin C, cetylpyridinium chloride and 2-hydroxypropyl-β-cyclodextrin. At this stage, the order in which the reagents are added is important.

At the second stage, the resulting complex is mixed with the excipients indicated in the composition, mixed by shaking and then pressed into tablets intended to obtain a solution for topical or external use.

Another object of the invention is the use of the claimed antibacterial composition as a local or external antibacterial agent, which consists in dissolving the antibacterial composition in water and applying the solution for the required purpose: rinsing the throat, wiping places that require disinfection, etc. The resulting solution can be used, for example, for tonsillitis, pharyngitis, tonsillitis, periodontal disease, gingivitis, stomatitis, etc.

The technical result of the invention consists in creating a new stable antibacterial composition for preparing a solution for local and external use based on the gramicidin C complex, which has improved solubility, which expands the arsenal of available antibacterial drugs for external use, in reducing the amount of excipients required by more than 3 times, in a new method for obtaining an antibacterial composition, in using an antibacterial composition in the form of a solution, which allows treating a significant area of the mucous membrane and external surfaces, without excessive salivation effect, more effectively cleaning to clean the oral cavity due to respiratory movements, the intensity of which is regulated by the patient himself, to minimize the toxic effect on the body and human microbiota due to the lack of ingestion of the solution into the body.

The stability of the claimed antibacterial composition was studied in the accelerated storage test, at a temperature of 40±2°C and a relative air humidity of 75±5% in three types of packaging: a strip of buffalo film (PAP/PE/ALU/PE), a strip of PET/F/PET/PE film, white blisters (PVC/ALU). It has been shown that the claimed pharmaceutical composition withstands this test for 6 months in buflen film (PAP/PE/ALU/PE) and PET/F/PET/PE film. without loss of active components, which corresponds to two years of storage under normal conditions (Table 2).

The antibacterial activity of the claimed antibacterial composition was studied in vitro . It was found that the claimed pharmaceutical has antimicrobial activity against all studied strains of Streptococcus pneumoniae and Staphylococcus aureus . The excipients in the composition did not prevent the release of the active ingredients of gramicidin C and cetylpyridinium chloride and did not affect their bactericidal action. The obtained values of the MIC and MBK, in the overwhelming majority of cases differing by no more than 4 times for different strains (Table 3), demonstrate the presence of a high bactericidal activity in the antibacterial composition against typical pathogens of the infectious and inflammatory pathology of the ENT organs and, in particular, the pathology of the oropharynx, which allows it to be effectively used in clinical practice for local and external use.

Appliances used:

- electronic laboratory scales AR 2140, AdventurerTM (Switzerland) and AUW-220D (Shimadzu, Japan);

- installation for obtaining purified water UVO-0.2 (Russia);

- Simplicity Water Purification System (Millipore, USA);

- climatic chamber SM 5/100-80 TVO (LLC "SM Climate", Russia);

- laboratory identifier of the disintegration process (ZAO Farmagen, Russia);

- desktop laboratory press Korsch type eko (Erweka, Germany);

- blister machine TOP BLISTER (Farmalabor Srl, Italy);

- automatic blister packing machine DPP-150 Al-plastic automatic blister packing machine, Zhejiang Jiangnan Pharmaceutical Machinery Co. (China).

- manual impulse apparatus CNT-200 (bag sealer) (CAS Corporation, Korea);

- high-pressure chromatograph LC-20 Prominence (Shimadzu, Japan) with a diode array detector and a Gemini C6-Phenyl column 4.6x250 mm (sorbent particle size 5 µm) (Phenomenex, USA) with a 3.0 mm guard column filled with the same sorbent;

- lyophilic dryer LS-1000; Prointekh LLC, Russia. Drying speed not less than 100 g/h. Condenser temperature -50°C, capacity 6 kg of ice. The working pressure in the vacuum chamber of the dryer is 6.67 Pa (5 10-2 mm Hg);

- Electrolux ENA38933W refrigerator, freezer compartment temperature -22°С.

The invention is illustrated by the following examples, but is not limited to them.

Example 1 Compositions of an antibacterial composition.

Composition 1.

gramicidin C - 3 mg;

cetylpyridinium chloride - 1 mg;

2-hydroxypropyl-β-cyclodextrin - 25 mg.

Composition 2.

gramicidin C dihydrochloride - 3.2 mg (in terms of gramicidin 3.0 mg);

cetylpyridinium chloride monohydrate - 1.05 mg (in terms of cetylpyridinium chloride 1.0 mg);

2-hydroxypropyl-β-cyclodextrin - 25 mg

Composition 3.

gramicidin C - 3 mg;

cetylpyridinium chloride - 1 mg;

2-hydroxypropyl-β-cyclodextrin - 25 mg.

mannitol - 256.75 mg;

crospovidone - 33 mg;

sorbitol - 32 mg;

aspartame (E951) - 8 mg;

flavor "green apple" - 8 mg;

flavoring "cinnamon" - 5 mg;

polyethylene glycol 4000 - 4 mg.

Composition 4.

gramicidin C - 3 mg;

cetylpyridinium chloride - 1 mg;

2-hydroxypropyl-β-cyclodextrin - 6 mg.

mannitol - 256.75 mg;

crospovidone - 33 mg;

sorbitol - 32 mg;

aspartame (E951) - 8 mg;

flavor "pear" - 6 mg;

polyethylene glycol 4000 - 4 mg.

Composition 5.

gramicidin C - 3 mg;

cetylpyridinium chloride - 1 mg;

2-hydroxypropyl-β-cyclodextrin - 100 mg.

mannitol - 256.75 mg;

crospovidone - 33 mg;

sorbitol - 32 mg;

aspartame (E951) - 8 mg;

flavor "green apple" - 8 mg;

flavoring "cinnamon" - 5 mg;

polyethylene glycol 4000 - 4 mg.

Composition 6.

gramicidin C - 3 mg;

cetylpyridinium chloride - 1 mg;

2-hydroxypropyl-β-cyclodextrin - 25 mg.

mannitol - 256.75 mg;

flavor "green apple" - 8 mg;

flavoring "cinnamon" - 5 mg.

Composition 7.

gramicidin C - 3 mg;

cetylpyridinium chloride - 1 mg;

2-hydroxypropyl-β-cyclodextrin - 25 mg.

mannitol - 260.0 mg;

crospovidone -35.0 mg;

aspartame - 10 mg

polyethylene glycol 4000 - 5.0 mg.

Composition 8.

gramicidin C - 3 mg;

cetylpyridinium chloride - 1 mg;

2-hydroxypropyl-β-cyclodextrin - 15 mg.

mannitol - 252 mg;

crospovidone - 33 mg;

sorbitol - 30 mg;

aspartame (E951) - 6 mg;

polyethylene glycol 4000 - 4 mg.

Composition 9.

gramicidin C - 3 mg;

cetylpyridinium chloride - 1 mg;

2-hydroxypropyl-β-cyclodextrin - 10 mg.

mannitol - 241.75 mg;

crospovidone - 33 mg;

sorbitol - 32 mg;

aspartame (E951) - 8 mg;

flavor "green apple" - 8 mg;

flavoring "cinnamon" - 5 mg;

polyethylene glycol 4000 - 4 mg.

Example 2. Obtaining the claimed antibacterial composition.

0.213 g of cetylpyridinium chloride is weighed on a balance and 5.088 g of 2-hydroxypropyl-β-cyclodextrin is placed in a 50 ml beaker, 7.3 ml of water is added. The glass is placed on a magnetic stirrer and stirred until a clear solution is obtained. Then, 0.639 g of gramicidin C is added to the resulting solution and stirred until a clear or slightly opalescent solution is obtained for 40-50 minutes. The resulting solution is transferred to a freezer and frozen at -22°C. The frozen solution is then freeze-dried. The resulting complex is crushed in a porcelain mortar and sieved through a sieve (0.5 mm). 53.71 g of briquetted mannitol are successively added to the resulting complex; 0.8356 g crushed and sieved PEG 4000; 6.897 g crospovidone; 6.688 g sorbitol; 1.6772 g of aspartame and "green apple" flavor and 1.045 g of "cinnamon" flavor, mixed by shaking for 5-20 minutes. The procedure will be similar for other ratios of components. Weight loss during screening is about 1%. The resulting composition is compressed into tablets to prepare a solution for topical and external use. Get a pharmaceutical composition corresponding to Composition 3 of Example 1. Comparison with the prototype is shown in table 1.

Compositions 1,2,4-9 were prepared in a similar way, adding the components of the pharmaceutical composition in the proportions indicated in Example 1 without changing the order of their mixing. For the preparation of each composition of cetylpyridinium chloride was taken in an amount of 0.2-0.3 g, gramicidin C - 3 times more by weight. The remaining components after the preparation of the antibacterial complex were added in accordance with the indicated compositions in Example 1.

The solubility of gramicidin C was determined as follows. A saturated solution was prepared from the antibacterial composition (Compositions 1-9 of Example 1) by adding an excess of gramicidin C and the claimed composition so that after 30 minutes after stirring a visible precipitate remained on a magnetic stirrer, the precipitate was filtered from the precipitate through a filter with a pore size of 0.22 μm, and the content of gramicidin C was quantitatively determined in the resulting solution by HPLC according to the method in accordance with the description of example 3. It was found that for different compositions of the compositions after their dissolution in water, the content of gramicidin C in it is 8 or more times higher in comparison with a solution of gramicidin C.

Variants of tablets with a weight of 370 mg or other weight have been obtained, which may be limited only by the available set of tablet press molds. Tablets disintegrated in less than 3 minutes in the disintegration test in accordance with SP XIV OFS.1.4.2.00013.15.

Example 3 Determination of the stability of an antibacterial composition.

In order to assess more distant chemical effects and to assess the impact on the stability of the drug formulation of short-term deviations from natural storage conditions (“accelerated aging”), experimental samples of tablets of the claimed antibacterial composition (20 tablets per point) were placed in three types of primary packaging for storage at a temperature of 40 ± 2 ° C and relative humidity of 75 ± 5%. Samples were analyzed after 22 days, 45 days, 3 and 6 months.

The test was carried out in accordance with the standard methodology “Stability and shelf life of medicines in accordance with the Global Fund XIV OFS.1.1.0009.18.

Quantitative analysis of the content of the components was performed on a Shimadzu high-pressure chromatograph with a diode array detector and a Gemini C6-Phenyl column 4.6 × 250 mm (sorbent particle size 5 μm) and a pre-column (3 mm) filled with the same sorbent (Phenomenex, USA) in a gradient elution mode with a mixture of 0.1 M solution of monosubstituted sodium phosphate with pH 3.0 (mobile phase A) and methane ol (mobile phase B), eluent flow rate 1 ml/min, dosed sample volume 20 μl, detection wavelength 220 nm; column thermostat room temperature.

The tablets of the antibacterial composition of the compositions 1-9 of Example 1 were packed in two types of packaging: the combined material Buflen (material composition: PAP/PE/ALU/PE); material combined PET/F/PET/PE.

The results of studying the stability of the antibacterial composition under conditions of "accelerated aging" in three types of packaging at a temperature of 40±2°C and a relative humidity of 75±5% are presented in Table 2 (Composition 3). The results obtained indicate the stability of the GLF in each of the packages. The content of active substances during accelerated storage of the antibacterial composition of Compositions 1, 2, 4-9 of Example 1 differed by no more than 10% from Composition 3.

Example 4. Determination of the antibacterial activity of the claimed antibacterial composition.

To confirm the antibacterial properties of the claimed antibacterial composition (Composition 3, Example 1), in comparison with gramicidin C and cetylpyridinium chloride, strains of microorganisms were used, which are the causative agents of ENT infections. One of the main causative agents of ENT infections are Streptococcus pneumoniae and Staphylococcus aureus . The study used strains of clinical isolates with characterized antibacterial resistance of various degrees of latitude:

Reference strain - Streptococcus pneumoniae ATCC 6303;

Clinical isolates - Streptococcus pneumoniae 711/58 serotype 4, Streptococcus pneumoniae 7670 serotype 13, Streptococcus pneumoniae 7779 serotype 13, Streptococcus pneumoniae 1068 serotype 19 A, Streptococcus pneumoniae 8074 serotype 6 ABCD;

Reference strain - Staphylococcus aureus ATCC 6538-P;

Clinical isolates - Staphylococcus aureus 758 (MRSA), Staphylococcus aureus 952 (MSSA), Staphylococcus aureus 402 (MRSA), Staphylococcus aureus 299 (MRSA), Staphylococcus aureus 176 (MRSA).

The determination of the antimicrobial activity of the tested objects was carried out by microdilution in tablets with the determination of the main quantitative indicator characterizing the microbiological activity of antibacterial drugs - the value of the minimum inhibitory concentration (MIC).

The prepared working solutions of the test objects (final concentration of the active substance from 256 µg/ml to 1 µg/ml) were added to 96-well plates. An inoculum of a microorganism strain at a given concentration was added to each well containing the required amount of the test object solution. As a positive control, wells containing a nutrient medium inoculated with a microorganism were used, as a negative control, wells with a nutrient medium were used.

After all agents were added, the plates were capped and incubated in a thermostat. 19 hours after incubation, the results were taken into account visually in transmitted light, with sufficient growth of the test microorganism, that is, with obvious clouding or color change of the nutrient medium. The minimum inhibitory concentration of the studied objects corresponded to the well with the lowest concentration of the studied objects without signs of visible growth of test microorganisms (the clear broth or the color of the medium did not change).

After determining the MIC, seedings were made from all wells without signs of visible growth on Petri dishes with a dense nutrient medium to determine the MBK. The minimum bactericidal concentration is the minimum concentration of an antibiotic in a nutrient medium at which its bactericidal effect is manifested in relation to the pathogen. For MBC take the concentration that ensures the death of 99.99% of the cells of the microbial population.

The results of determining the antibacterial activity are presented in Table 3. Based on the results of the study, it was found that the pharmaceutical composition has antimicrobial activity against all the studied strains of Streptococcus pneumoniae and Staphylococcus aureus . The presence of excipients in the antibacterial composition did not prevent the release of the active substances Gramicidin C and Cetylpyridinium and did not affect their bactericidal action. The obtained values of MIC and MBK, in the overwhelming majority of cases differing by no more than 4 times, demonstrate the presence of a high bactericidal activity in the antibacterial composition against typical pathogens of the infectious and inflammatory pathology of the ENT organs and, in particular, the pathology of the oropharynx, which allows it to be effectively used in clinical practice.

Table 1. The composition of the excipients of the claimed antibacterial composition and comparison with the prototype No. The claimed pharmaceutical composition (composition 3) Weight, mg Prototype "Grammidin ^® neo" Weight, mg 1 Mannitol 256.75 silicon dioxide colloidal 5 2 Crospovidone 33 talc 60 3 Sorbitol 32 acesulfame potassium 15 4 2-hydroxypropyl-β-cyclodextrin 25 mint flavor 25 5 aspartame (E951) 8 sorbitol 1065.75 6 green apple flavor 8 magnesium stearate 25 7 flavoring "cinnamon" 5 8 polyethylene glycol 4000 4

Table 2 Stability data of the antibacterial composition (Formulation 3 Example 1) in the accelerated storage test (40 ^° C., 75% humidity). Type of packaging 1 - strip of buffalo film (PAP/PE/ALU/PE); 2 - PET/F/PET/PE film strip Type of packaging Shelf life, days 0 22 45 90 180 The content of active substances, the initial values of gramicidin C - 3 mg; cetylpyridinium chloride - 1 mg; deviation of values ±10% is allowed 1 2.94±0.05
1.14±0.02 3.12±0.07
1.11±0.03 3.07±0.02
0.96±0.01 3.10±0.01
1.09±0.03 2.99±0.06
1.15±0.03 2 2.94±0.05
1.14±0.02 3.01±0.04
1.04±0.02 3.09±0.05
0.98±0.02 3.07±0.09
1.08±0.05 3.04±0.01
1.15±0.00

Table 3. Antibacterial activity of the claimed antibacterial composition in comparison with gramicidin C and cetylpyridinium chloride No. p / p Test
microorganism Gramicidin C Cetylpyridinium chloride pharmaceutical composition IPC MBK IPC MBK IPC MBK Streptococcus pneumoniae ATCC 6303 4 8 8 8 4 4 Streptococcus pneumonia 711/58 serotype 4 16 16 16 32 8 16 Streptococcus pneumoniae 1068 4 4 16 16 4 4 Streptococcus pneumonia 7670 4 4 8 16 4 4 Streptococcus pneumoniae 7779 4 4 8 16 4 4 Streptococcus pneumonia 8074 4 4 8 16 4 4 Staphylococcus aureus ATCC 6538-P 2 2 2 2 2 2 Staphylococcus aureus 176 4 8 2 8 4 8 Staphylococcus aureus 299 4 8 4 4 4 8 Staphylococcus aureus 402 2 8 2 4 2 4 Staphylococcus aureus 758 2 2 2 4 2 2 Staphylococcus aureus 952 2 8 2 2 2 4

Claims (12)

1. Antibacterial complex, characterized in that it consists of 2-hydroxypropyl-β-cyclodextrin, gramicidin C or its pharmaceutically acceptable salt (in terms of gramicidin C), cetylpyridinium chloride or its monohydrate (in terms of cetylpyridinium chloride) in a mass ratio of 6-100:3:1. 2. The antibacterial complex according to claim 1, characterized in that the mass ratio of the components is 25:3:1. 3. An antibacterial composition for preparing a solution for local or external use, characterized in that it is made in the form of a tablet containing the antibacterial complex according to claim 1 as an active principle and mannitol, crospovidone, sorbitol, aspartame, polyethylene glycol 4000 as auxiliary components with the following content of components: antibacterial complex - 10.0-104.0 mg; mannitol - 230.0-270.0 mg; crospovidone - 30.0-36.0 mg; sorbitol - 29.0-35.0 mg; aspartame - 6.0-10.0 mg; polyethylene glycol 4000 - 3.0-5.0 mg. 4. An antibacterial composition according to claim 3, characterized in that it additionally contains at least one flavor as excipients. 5. A method for producing an antibacterial composition for preparing a solution for local or external use according to claim 3 or 4, characterized in that an antibacterial complex according to claim 1 is obtained by dissolving cetylpyridinium chloride or its monohydrate and 2-hydroxypropyl-β-cyclodextrin in water, stirring until a clear solution, adding gramicidin C or its pharmaceutically acceptable salt to the solution, stirring for 40-50 minutes until a transparent or slightly opalescent solution, freezing the solution, freeze-drying, followed by the addition of excipients to the resulting complex, mixing and tableting. 6. The use of the composition according to claim 3 for obtaining an antibacterial agent of local or external action in the form of an aqueous solution.