RU2595852C1 - Antibacterial suppositories - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to suppositories for treating and preventing gynecological diseases. Offered antimicrobial suppositories contain poly-chloride (4,9-dioxadodecane guanidine), copper hydrocolloid, Vitepsol, cocoa oil, glycerin and polyethylene glycol.

EFFECT: invention provides increasing antimicrobial action with simultaneous absence of toxicity of the drug in total.

1 cl, 2 tbl, 3 ex

Description

The invention relates to medicine, in particular to vaginal suppositories of antimicrobial action, which can be used to treat diseases of the female genital area.

Prevention and treatment of gynecological diseases is an urgent problem. These diseases - bacterial vaginosis, candida vomit and Trichomonas colpitis - require the use of targeted drugs. Often the infection is mixed, which requires combined drugs or broad-spectrum antiseptics.

Known suppositories of anti-inflammatory, antiseptic and regenerative effects, including in their composition the following components, wt. %:

1,3-diethylbenzimidazolium triiodide 1.5-3.0 Polyvinylpyrrolidone 6-12 Hydrophobic, lipophilic base Rest

(RF patent No. 2246945, class A61K 9/02, 2005).

The disadvantages of the known funds include the irritating effect of its use and low therapeutic activity.

The closest in combination of essential features composition for the same purpose are suppositories according to UK patent No. 2163649, 1984.

The known composition of antifungal suppositories for the treatment of vaginal infections contains an antifungal substance, in particular, nystatin, hydrocolloid - sodium carboxymethyl cellulose or hydroxypropyl methyl cellulose and a hydrophobic base with a low melting point - a mixture of Vitepsol W-35 - (31.2%) and Vitepsol N-15 ,8%). Moreover, these components are taken in the following ratio, mg / 1 suppository:

Nystatin (micropulverized) (5000 U / mg) 5-50 Hydrocolloid (sodium carboxymethyl cellulose or hydroxypropyl methylcellulose) 10-50

Low melting base

(Vitepsol W-35 - (31.2%), Vitepsol N-15 (68.8%)) 985-900

Known suppositories after introduction into the vagina are easily melted. When melting due to exposure to body temperature, hydrocolloid (hydroxypropyl methylcellulose) and fungicidal agent are released and, due to improved adhesion to the vaginal mucosa, the antifungal substance is evenly distributed at the site of infection. However, the introduction of, in particular, sodium carboxymethyl cellulose into a hydrophobic base does not provide a uniform distribution of the drug substance throughout the volume of the suppository mass, which affects the quality of the target product. In addition, nystatin is easily destroyed in an acidic and alkaline environment, is thermally unstable and is oxidized by atmospheric oxygen. This leads to a deterioration in the quality of suppositories and a reduction in their shelf life. It should be borne in mind that nystatin, being an antibiotic, contributes to the appearance of allergic reactions and the body’s addiction to antibiotic treatment.

The objective of the invention, which determines its purpose, is to expand the spectrum of therapeutic activity and increase consumer characteristics.

To solve the technical problem in antimicrobial suppositories, including an antimicrobial agent, a hydrocolloid agent, and Vitepsol, poly- (4,9-dioxadodecanguanidine) chloride is used as an antimicrobial agent, copper hydrocolloid is used as a hydrocolloid agent, and cocoa butter, glycerol and polyethylene are added the following ratio of components in g per suppository weighing 2.0 g:

Poly- (4,9-dioxadodecanguanidine) chloride 0.01-0.04 Cacao butter 0.03-0.07 Glycerol 0.02-0.06 Polyethylene glycol 0.01-0.04 Copper hydrocolloid 0.004-0.1 Vitepsol Rest

The invention can be explained as follows.

Poly- (4,9-dioxadodecanguanidine) chloride (PDDG chloride) - is introduced into the composition as an antimicrobial agent. PDDG chloride has a wide spectrum of antimicrobial activity and is a new generation biocide. According to its characteristics, PDDH chloride is a low-toxic compound (IV class of danger) and does not cause allergies. PDDG chloride is produced as a substance under the ECOSEPT trademark by the International Institute of Ecological and Technological Problems (LLC MIETP, Russia).

The effect of PDDH chloride on microorganisms can be represented as follows.

- Guanidine polycations are adsorbed on the negatively charged surface of the bacterial cell and block its nutrition.

- PDMD chloride macromolecules diffuse through the cell wall and cause its irreversible structural damage.

- PDDH chloride binds to acid phospholipids, proteins of the cytoplasmic membrane, resulting in blockade of glycolytic enzymes of the respiratory system, loss of pathogenic properties and death of microbial cells.

Cocoa butter is used in suppositories for more efficient transportation and resorption of the active therapeutic components of the suppository in the mucous membranes.

Cocoa butter is a dense-bodied vegetable fat derived from chocolate tree seeds.

Cocoa butter at room temperature has a light yellow color, a slight aromatic odor. Cocoa butter manufactured by ICAM S.p.A (Italy) was used in the invention.

Cocoa butter contains trace elements and vitamins, anti-inflammatory components and acts as an antioxidant. The healing and tonic properties of cocoa butter are provided by the presence of caffeine, tannin and methylxanthine in the composition.

Glycerin is introduced into the composition of suppositories as a dispersant, ensuring a uniform distribution of antimicrobial agents.

Its use allows you to get the optimal therapeutic effect.

Glycerin is a transparent viscous liquid with a basic substance content of at least 99 wt. % (GOST 6259-75).

Polyethylene glycol is a polymer compound of ethylene glycol with various degrees of polymerization.

Structural formula:

Polyethylene glycol in suppositories acts as a dispersant and plasticizer, promotes a uniform distribution of medicinal substances, helps to increase the stability of the composition of suppositories, and provides a slow release of drugs during use.

Polyethylene glycol of brands PEG-600, PEG -1500, PEG-6000 manufactured by BASF of the Macrogol brand (Germany) is used.

As a hydrocolloid agent, a copper hydrocolloid is introduced into the suppository.

The process of synthesizing copper hydrocolloid was carried out at YurDex-Eco LLC.

The process itself consists in obtaining germ cells of a hydrocolloid and the synthesis of copper hydrocolloid proper with a given particle size.

Sodium borohydride can act as a reducing agent; the general recovery scheme is presented below:

67 ml of distilled water are added to a 250 ml flask and 0.4 g of stabilizer is dissolved. The stabilizer may be polyvinyl alcohol, polyvinylpyrrolidone or polymer derivatives of guanidine. In this synthesis, poly- (4,9-dioxadodecanguanidine) chloride was used as a stabilizer. The mixture is stirred while heating to 40-45 ° C, after which a solution of 0.3 g of copper pentahydrate in 23 ml of distilled water is added to the resulting solution, stirred until complete dissolution and acquisition of a solution of opalescent bluish color.

After complete dissolution of the components, a solution of 0.04 g of reducing agent in 10 ml of distilled water is added to the flask. As a reducing agent, glucose or sodium borohydride can be used. In this synthesis, sodium borohydride was used as a reducing agent. Next, the system is heated to 50-54 ° C and, with stirring, dry NaOH is added in portions of ~ 0.015 g, while the pH of the system should be kept at 10.8-11.2. The addition of portions of caustic soda is stopped after acquiring a solution of yellow-orange color, gradually turning into a saturated reddish-orange, after which the system is stirred for 30-90 minutes with slow cooling.

Copper hydrocolloid is a yellow to light brown colloidal solution with a copper concentration of 0.007-0.016 wt. %, while the average particle size of the copper hydrocolloid obtained by this method is 18-20 nm with an absorption maximum of 419 nm.

It is known that copper and its compounds are used in medicine as an antiseptic component that has anti-inflammatory bactericidal, antimicrobial and astringent effects.

Various sources have shown that copper hydrocolloid particles, on the one hand, have very low toxicity, and, on the other hand, exhibit a high antimicrobial effect with respect to cells of test cultures of gram-positive and gram-negative bacteria, which creates the prerequisites for using them as part of wound healing preparations.

In addition, copper and its compounds take part in reparative processes and cell division processes (Babushkina I.V., Gladkova E.V., Mamonova I.A., Norkin I.A., Puchinyan D.M. Biological activity of copper nanoparticles in experiment // Russian Scientific World No. 6, pp. 1204-1207, 2014; Maqusood Ahamed, Hisham A. Alhadlaq, M.A. Majeed Khan, Ponmu-rugan Karuppiah and Naif A. Al-Dhabi. Synthesis, Characterization, and Antimicrobial Activity of Copper Oxide Nanoparticles // Volume 2014 (2014). Article ID 637858. 4 p).

The joint introduction of copper hydrocolloid and poly- (4.9 dioxadodecanguanidine) chloride into the composition of the proposed suppositories provides an increasing antimicrobial effect with the simultaneous absence of toxicity of the drug as a whole.

Vitepsol brand H-15 and W-35 as a suppository base promotes a uniform distribution of active components and their good bioavailability from the base.

The base, consisting of Vitepsol brand N-15 and W-35, increases the shelf life of suppositories, provides good consistency and hardness, which improves their presentation and creates ease of use.

This base also contributes to a sharp transition from solid to liquid, bypassing the stage of softening. Use Vitepsol brand H-15 and Vitepsol brand W-35 from Cremer Oleo GmbH & Co. KG (Germany) or according to VTU No. 3-95.

The number of components of the composition of the suppository is the optimal value and is selected on the basis of numerous experiments.

The developed optimal combination of components leads to a homogeneous composition capable of distributing the drug throughout the cavity after administration of the suppository.

The content of antibacterial components is strictly regulated.

The proof of the optimality of the selected content limits of the suppository components is given in table 1, which also shows the properties of suppositories made of components, the number of which went beyond the stated content limits.

Also, confirmation of the optimal number of components of the proposed suppository in the selected intervals of the ratio of the components compared to suppositories made of components whose content was outside the selected limits was confirmed by studies on the bioavailability of the active components of the suppository.

The determination of comparative bioavailability was carried out according to the method (Muravyov I.A. Technology of drugs. - M., 1980, v. 1). Installation for diffuse release is a glass tube with a length of 15 cm, a cross-sectional area of 10 cm ² , one end of which with a fixed membrane is lowered to a depth of 2-3 cm in Ringer's solution. After reaching a temperature of 37 ° C, a suppository was placed on the membrane and samples were taken using a pipette at regular intervals. Analysis of samples taken every 15 minutes showed that the release of active antimicrobial components from suppositories obtained in the proposed intervals for the content of the components of the suppository base is faster and, therefore, the composition of the proposed suppositories is optimal.

Suppositories are prepared as follows.

At the first stage, the basis is prepared. To do this, the calculated amount of Vitepsol grade N-15 and grade W-35 is loaded into a heated reactor with stirring and melted at a temperature of 60-65 ° C. In another container, cocoa butter is separately melted at a temperature of 55-60 ° C, then pumped into the main reactor, melted at a temperature of 60-65 ° C and mixed until a homogeneous mass is obtained. After the mass cools down to 45-48 ° C, polyethylene glycol, glycerin, poly- (4,9-dioxadodecanguanidine) chloride, copper hydrocolloid are successively added to the reactor with stirring, and all components are mixed until a homogeneous mass is obtained. If necessary, carry out additional grinding of the powder ingredients in a ball mill. The temperature in the reactor is maintained up to the pouring stage. The product in the reactor remains in a molten state at a constant temperature of 45-48 ° C. Next, the molten mixture is fed to the molding tape and spilled into the molding containers. After molding, the suppositories enter the cooling unit. At the last stage after cooling, it is necessary to carry out sealing, cutting and labeling of suppository containers.

It should be noted that the resulting suppositories can have a mass of from 1.2 to 2.5 g, depending on the technological conditions of production.

The invention can be illustrated by the following examples of suppositories with a ratio of components in g per suppository weighing 2.0 g:

Example 1

PDDG Chloride 0.01 Cacao butter 0,07 Glycerol 0.05 Polyethylene glycol PEG-600 0,03 Copper hydrocolloid 0.004 Witepsol W-35 1,836

Example 2

PDDG Chloride 0,03 Cacao butter 0.05 Glycerol 0.02 Polyethylene glycol PEG-1500 0.04 Copper hydrocolloid 0.1 Witepsol W-35 1.76

Example 3

PDDG Chloride 0.04 Cacao butter 0,03 Glycerol 0.06 Polyethylene glycol PEG-6000 0.01 Copper hydrocolloid 0.05

A mixture of Vitepsol H-15 and W-35, respectively

H-15 - 0.45 g, W-35 - 1.46 g 1.91

The remaining examples are summarized in table 1. For comparison with the characteristics of the prototype was made suppository of the following composition in g per suppository weighing 2.0 g:

Example (prototype)

Nystatin (5000 IU / mg) 0.04 Hydrocolloid - sodium carboxymethyl cellulose 0,03

The basis of the mixture of Vitepsol H-15 and W-35, respectively

Vitepsol N-15 - 1.33 g, Vitepsol W-35 - 0.60 g, 1.93

To prepare suppositories of known composition, the base is heated to a temperature of not more than 45 ° C until the base is completely melted. Then the temperature of the mass is reduced to 40 ° C and a hydrocolloid, in particular sodium carboxymethyl cellulose, is introduced with stirring. After this, an antifungal substance, in particular nystatin, is added and stirring is continued until a relatively uniform suspension is formed. After this, the stirring is completed, the mass is poured into molds, so that upon cooling to obtain suppositories.

The therapeutic efficacy of known vaginal suppositories with nystatin is achieved by introducing a hydrocolloid in them - sodium carboxymethyl cellulose or hydroxypropyl methyl cellulose.

The suppositories of the invention have been tested for cytotoxicity in a NIH / 3T3 mouse fibroblast culture at the accreditation center of the Institute of Biomedical Tests and Technologies. The extracts from the presented samples of candles were investigated (according to examples 1, 2 and 3). The extracts were filtered through a filter nozzle for a syringe with a pore diameter of 0.22 μm. The extraction time was 4 hours. During the extraction process, the candles dissolved. The hood was an opaque liquid with fat droplets.

The tests showed that the extracts from the presented samples of suppositories (according to examples 1, 2 and 3) do not exert cytotoxicity on the culture of mouse fibroblasts NIH / 3T3.

The antimicrobial activity was checked according to the Guidelines for preparations containing antimicrobial components according to the method of agar plates.

Müller-Hinton agar was used as a solid nutrient medium, which was contaminated with a microbial suspension of daily test cultures (in an amount of 10 ⁵ CFU per 1 ml). The main clinical microorganism strains were used as test cultures: Staphylococcus epidermidis, Staphylococcus aureus, Esherichia. Coli, Enterococcus spp., Candida albicans, Pseudomonas aeruginosa and Bacillus subtilis.

A 0.8 cm diameter well was made on the surface of contaminated agar poured into Petri dishes. The studied suppository samples were placed in the well (according to examples 1, 2, and 3, as well as a suppository sample made according to the prototype), after which the crops were incubated in an incubator with 37 ° C for 24 hours

The antimicrobial activity of the test material was evaluated by the degree of inhibition of microbial growth, as well as the size of the zone of growth inhibition of microorganisms from the edges of the well, expressed in millimeters. The test results are presented in table 2.

According to the results, the proposed suppositories had a high antimicrobial activity against gram-positive microflora (Staphylococcus epidermidis, Staphylococcus aureus and Enterococcus spp), gram-positive spore-forming Bacillus subtilis, Escherichia coli (Esherichia. Coli), in relation to fungus Candida albidae , as well as gram-negative microflora (Pseudomonas aeruginosa).

The growth retardation zone of Staphylococcus epidermidis and Staphylococcus aureus microflora for suppositories (according to examples 1, 2 and 3) was 24–28 mm and 18–20 mm, respectively, which indicates a high antimicrobial activity of suppositories, because It is known that infections of the genitourinary system caused by Staphylococcus epidermidis and Staphylococcus aureus can be very diverse, for example, inflammatory diseases, cystitis, pyelonephritis, vulvovaginitis, urethritis. With the ascending penetration of the pathogen, more serious diseases develop, such as endometritis, prostatitis, interstitial nephritis, etc.

Enterococcus spp and Esherichia are known. Coli are one of the causative agents of inflammatory diseases of the female genital area.

The growth retardation zone of Enterococcus spp microflora for suppositories (according to examples 1, 2 and 3) was 14-18 mm, and the growth retardation zone of Esherichia microflora. Coli was 18-22 mm, which is also a high estimate of the antimicrobial activity of the presented candle samples.

It is known that Pseudomonas aeruginosa, the causative agent of urinary tract infections, is characterized by significant natural resistance to most antimicrobial agents used in clinics, which causes severe complications after infection of the genitourinary system.

Tests showed that the zone of growth inhibition of microflora according to Pseudomonas aeruginosa for suppositories (according to examples 1, 2 and 3) was 14-16 mm.

The zone of growth inhibition of microflora according to Candida albicans for suppositories (according to examples 1, 2 and 3) was 16-22 mm. It is known that one of the main targets for defeat by Candida fungi is the vaginal mucosa, which is the cause, including urogenital candidiasis, which is caused by Candida albicans in 80-90%.

Suppositories made according to the prototype showed lower antimicrobial activity (table 2).

The proposed suppositories due to their high effectiveness are recommended for wide medical use.

The drug does not develop the resistance of microorganisms. After the use of suppositories, the normal vaginal microflora is quickly restored. In addition, a prolonged effect of therapeutic components is achieved throughout the infected surface.

Suppositories can be used to treat vaginitis (nonspecific and mixed infections) of bacterial vaginosis, infections caused by fungi of the genus Candida, trichomoniasis, as well as for the prevention of infectious complications during gynecological and diagnostic procedures.

The proposed antimicrobial suppositories meet the requirements for suppositories, namely:

- possess optimal rheological indicators, corresponding structural and mechanical properties and provide maximum contact between medicinal substances and the mucous membrane;

- do not have an irritating effect;

- retain high antimicrobial activity during prolonged storage;

- easily release medicinal substances, contribute to the manifestation of the pharmacological action of medicinal substances.

Claims (1)

Antimicrobial suppositories, including an antimicrobial agent, a hydrocolloid agent and Vitepsol, characterized in that poly- (4,9-dioxadodecanguanidine) chloride is used as an antimicrobial agent, copper hydrocolloid is used as a hydrocolloid agent and additionally contains cocoa butter, glycerin and the following polyethylene glycol the ratio of components in g per suppository weighing 2.0 g:
Poly- (4,9-dioxadodecanguanidine) chloride 0.01-0.04 Cacao butter 0.03-0.07 Glycerol 0.02-0.06 Polyethylene glycol 0.01-0.04 Copper hydrocolloid 0.004-0.1 Vitepsol Rest