RU2356535C2 - Synergistic suppositories for infectious gynecopathies treatment (versions) - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to pharmaceutics and concerns suppositories for infectious gynecopathies treatment containing nitroimidazole compounds as an antibacterial and antimicrobial agent, polyene macrolide compounds as an antifungal agent, and comfort-improving local anesthetic in diphilic or hydrophilic suppository base, as well as alpha or beta amino acid reparative agent as an additive ensuring accelerated healing of infected tissues.

EFFECT: suppositories ensure improved clinical effectiveness.

12 cl, 2 dwg, 10 tbl, 17 ex

Description

FIELD OF TECHNOLOGY

The invention relates to biocidal drugs for external use, in particular to dosage forms with an elastic-viscoplastic medium intended for the treatment of infectious gynecological diseases.

BACKGROUND

Currently, due to the high frequency of protozoan, fungal and bacterial infections of the lower part of the female genital organs, topical biocidal preparations are of great importance in gynecology. In view of the great variability and wide prevalence of strains of the causative agents of vulvitis and vaginitis, the breadth of the spectrum of action of the drug used is of particular importance [A. Krasilnikov Handbook of antiseptics. - Minsk: Higher School, 1995. - 367 p.].

The use of an effective composition of substances with antiprotozoal, antimicrobial and fungicidal action is only part of the solution, the rest is left to the creation of a rational dosage form. In most cases, there is a need to ensure uniform distribution of the drug over the entire surface of the vaginal mucosa to achieve total destruction of the pathogen, as well as a prolonged action. The intravaginal dosage form should provide comfort during administration (not to injure the mucous membrane, have an optimal consistency and time of complete deformation / dissolution) and high bioavailability of the active substances [Industrial technology of drugs / ed. prof. V.I. Chueshov - Kharkov: NFAA, - 2002].

Among the prolonged dosage forms of local action used in gynecology, tablets, ointments, suppositories can be listed. Soft dosage forms (suppositories and ointments) have an advantage over aqueous solutions, providing a more significant prolongation of the therapeutic effect due to the high viscosity and structure [Sinev DN, Marchenko LG, Sineva TD Reference manual for pharmacy technology of drugs. - St. Petersburg: Nevsky dialect, 2001. - 316 p.]. The advantages of hydrophilic and diphilic suppository bases over lipophilic ones are known: the best solubility in the vaginal secretion, distribution over the mucous membrane, release parameters, reduced risk of microbial contamination [Polymers in Pharmacy / ed. A.I. Tenzova, Medicine, Moscow (1985)].

The diphilic bases can be represented by the products of directed esterification and hydrogenation of fatty acids (vitepsol, massupol, lazupol), as well as mixtures of surface-active (Twins, Spens, poloxamers, emulsifiers), lipophilic (fats, waxes) and hydrophilic substances. Polyethylene glycols (macrogols), gelatin gels, and higher fatty alcohols are used as hydrophilic bases.

Known compositions of vaginal suppositories intended for the treatment of bacterial vaginitis and containing, as the main active substance, antibacterial and antimicrobial agents, preparations of a number of nitroimidazoles (e.g. metronidazole), and additionally optionally containing an antibiotic as an active substance of antifungal action. A common drawback of these solutions is the poor solubility of the active components in lipophilic, diphilic and hydrophilic suppository bases. This phenomenon directly affects the release of drugs from the base and the implementation of the therapeutic effect.

Among the vaginal suppositories of antifungal action is known the drug "Pimafucin" (state registration number P No. 013552 / 01-2002), manufactured by Yamanouchi, which contains the polyene macrolide antifungal agent natamycin (pimafucin) as the active substance. The advantage of introducing natamycin into the suppository is its low toxicity, it is indicated for administration to pregnant women [Vidal Handbook. Medicines in Russia: A Handbook. B-476, publishing house ZAO AstraFarmService, 2002].

US application 20060241061 states that natamycin is slightly soluble in ethanol and water, but raising the pH to 12 with sodium hydroxide increases solubility. Such a change in pH can adversely affect the state of the vaginal tract, which normally has a pH of 4-5.

According to European patent No. 1642500, natamycin is less active in crystalline form than in solution. This fact may indicate the introduction of a solvent for natamycin in the prescription.

Russian patent No. 2032402 describes compositions for administration to the vagina, while the pH of the cream, according to the authors of the patent, is optimized for the normal pH of the vagina and is 3.75-4.75. In addition, a number of surfactants, preservatives and thickeners are given in the patent description. Ethanol and other monohydric alcohols, as well as propylene glycol and other polyhydric alcohols are given as adjuvants and cosolvents.

In a number of patents, propylene glycol is used as an excipient in an amount of preferably not more than 5 wt.%. Russian patent No. 2032402 contains metronidazole in a mixture of polyethylene glycols, which are dissolved in vaginal secretion. In US application No. 20050080038, a composition of metronidazole and propylene glycol in the form of a vaginal gel is indicated, in US application No.2006024243 a composition of metronidazole with propylene glycol in the form of a foam, in the international application WO 2006/050303, a buffered composition for treating bacterial vaginitis containing metronidazole in combination with .

However, in none of the compositions the optimal dispersion of metronidazole and natamycin is achieved if they are used together with a prolonged effect.

Closest to our proposed solution is considered in US patent No. 6432935, which describes a pharmaceutical suppository composition consisting of an antibiotic, sulfonamide, nystatin and metronidazole. PEG and cocoa butter are used as the base.

The disadvantages of this solution is the low dispersion of the drugs used, due to the properties of the suppository base, which leads to a deterioration in the parameters of their release and delivery to the focus of infection, as well as low comfort of use and the possibility of abrasive effects on the vaginal mucosa.

In Russian patent No. 2195281 a composition of pessaries containing metronidazole, miconazole and lidocaine is claimed. Such a composition has a high comfort of use due to a local anesthetic, however, there are a number of disadvantages due to the presence of the azole antifungal agent miconazole: various manifestations of toxicity [T.S. Meeker, M.S. Siegel, F.M.Shiota, J.J. Crowley, R.W. Mcguffin. Toxicity of Amphotericin B, Miconazole, and Ketoconazole to Human Granulocyte Progenitor Cells In Vitro / Antimicrobial agents and chemotherapy, 1983, p. 169-171], allergenicity, presence of resistant forms of fungi.

DESCRIPTION OF THE INVENTION

The present invention is the creation of combined biocidal topical formulations for the treatment of a wide range of infectious diseases of the lower female genital organs, providing improved release of metronidazole and natamycin when used together with a prolonged action and comfort.

In accordance with the invention, intravaginal suppositories for the treatment of infectious diseases of the lower female genital organs are described containing, as an antibacterial and antimicrobial agent, a compound from a number of nitroimidazoles, polyene macrolide compounds and a local anesthetic in a diphilic or hydrophilic suppository base in the following ratio of components as an antifungal agent % wt .:

compound from a number of nitroimidazoles 15.0-30.0 macrolide polyene compound 4.0-6.0 local anesthetic 0.01-5.0 diphilic or hydrophilic suppository base rest

The following aspect of the present invention describes intravaginal suppositories for the treatment of infectious diseases of the lower female genital organs, containing, in addition to combining a number of nitroimidazoles as an antibacterial and antimicrobial agent, combining a number of polyene macrolides as an antifungal agent and local anesthetic, a reparative agent that promotes accelerated wound healing , in the following ratio of components,% wt .:

compound from a number of nitroimidazoles 15.0-30.0 macrolide polyene compound 4.0-6.0 local anesthetic 0.01-5.0 reparative agent 0.01-5.0 diphilic or hydrophilic suppository base rest

The following aspect of the present invention describes intravaginal suppositories for the treatment of infectious diseases of the lower part of the female genital organs, additionally containing a buffer system to maintain a natural pH level for the vagina, in the following ratio, wt.%:

compound from a number of nitroimidazoles 15.0-30.0 macrolide polyene compound 4.0-6.0 local anesthetic 0.01-5.0 reparative agent 0.01-5.0 buffer system 0.01-1.0 diphilic or hydrophilic suppository base rest

The following aspect of the present invention describes intravaginal suppositories for the treatment of infectious diseases of the lower female genital organs, additionally containing an antioxidant in the following ratio, wt.%:

compound from a number of nitroimidazoles 20.0-27.0 macrolide polyene compound 4.0-6.0 local anesthetic 0.01-5.0 reparative agent 0.01-5.0 antioxidant 0.01-2.0 diphilic or hydrophilic suppository base rest

In accordance with the present invention, the biocidal agents used in the nitroimidazole series are selected from the group consisting of metronidazole, tinidazole, secnidazole, ornidazole and the group of polyene antibiotics: natamycin, nystatin, amphotericin, rifamycin, are included in the current edition of the British, European, and American Pharmacopoeias.

In addition, local antibiotics are introduced into the claimed composition to increase comfort: lidocaine hydrochloride, xylocaine, bupivacaine, novocaine, benzocaine, which have a local analgesic effect. To accelerate the healing of tissues affected by infection, amino acids that accelerate repair (repair agents) are included in the composition of suppositories. In a preferred embodiment, the acids are selected from the group consisting of taurine, arginine, tryptophan, lysine, proline, histidine [Jacobsen, J .; Smith, L .; Biochemistry and Physiology of Taurine and Taurine Derivatives. Phys. Rev. 48; 1968], also included in the current editions of the American and European Pharmacopoeia.

In one embodiment of the invention, a buffering system is included in the suppository as a component for adjusting the pH. In a preferred embodiment of the invention, under buffer systems added to the composition for adjusting the pH value are meant ascorbate, carbonate, phosphate, citrate, oxalate buffers. In a preferred embodiment, 0.1 to 1 wt.% Ascorbate buffer is added to the suppository. The ascorbate buffer, in addition to adjusting the pH in the suppository, can act as an antioxidant to prevent the oxidation of active substances (for example, natamycin) during the preparation and packaging of the suppository.

According to another embodiment of the invention, compounds are added to the suppository as additional antioxidants whose antioxidant properties appear in diphilic and hydrophilic suppository bases. Such antioxidants are selected from the group consisting of ascorbic acid, ascorbyl palmitate, hypophosphoric acid, sodium ascorbate, vitamins C and E, sodium bisulfite, EDTA, citric acid and other compounds known to those skilled in the art that exhibit their antioxidant properties in polymeric diphilic or hydrophilic bases.

The combination and ratio of these components was established experimentally and is optimal according to the results of physico-chemical, pharmacotechnological and biological studies.

The dosage of metronidazole in vaginal suppositories (Flagil produced by Laboratories Aventis) is from 200 to 500 mg with a single dose of 1 g.

The dosage of natamycin in the preparations "Pimafucin" manufactured by Yamanouchi is 100 mg.

Lidocaine hydrochloride is taken in the minimum amount recommended in the Mashkovsky Handbook M.D. (Mashkovsky M.D. Medicines. - Kharkov, 1997) for local use in gynecology.

In accordance with the invention, the diphilic suppository base is a mixture of fat-soluble components (solid fat of various types, waxes) and water-soluble components (emulsifier No. 1, emulsifier T-2, Twins, Spans, glycols, distilled monoglycerins). In a preferred embodiment, solid fats are used as the fat-soluble components, and the emulsifier T-2, Tween-80, PEG-1500 (polyethylene glycol-1500) and propylene glycol are used as water-soluble components. In the most preferred embodiment of the invention, a diphilic suppository base is used in the following ratio, wt.%:

Type A solid fat 55.0-80.0 emulsifier T-2 4,5-6,0 Twin 80 1.5-3.0 PEG-1500 0.1-5.0 propylene glycol 0.1-10.0

In accordance with the invention, the components of the hydrophilic suppository base are water-soluble components: glycerins, glycols. In a preferred embodiment of the invention, PEG-1500 and propylene glycol are used as components of the hydrophilic base. In a most preferred embodiment of the invention, these components are included in a suppository base in the following proportions, wt.%:

PEG-1500 60.0-85.0 propylene glycol 0.1-20.0

Suppositories in accordance with the invention are characterized by the following physicochemical properties.

Melting point or period of complete deformation. For the developed compositions, the values of these characteristics correspond to the requirements of the State Pharmacopoeia (GF-XI), namely, the melting temperature does not exceed 37 ° C, and the time of complete deformation is less than 1 hour.

Dissolution time. The dissolution time of the suppositories according to the invention, and more precisely the time for the suppository to be transformed into an aqueous dispersion into a fine dispersion, meets the requirements set forth in the State Pharmacopoeia. Therefore, the compositions described in the invention provide an effective distribution of active substances in the mucous membrane.

pH The pH range of formulations with a diphilic base and a hydrophilic base is 5-7. The active substance - a compound from a number of nitroimidazoles - increases the pH of the suppository base. The pH value of the developed drugs corresponds to the physiological pH of the mucous membranes of the body and slightly exceeds the natural pH of the vagina (4-5). However, given the fact that the drug is used in inflammatory processes, acidic pH values may be undesirable.

Structural and mechanical properties. The determination of the optimal consistency of suppositories was carried out by measuring the dynamic viscosity of the developed formulations (see table 5 in the "Examples" section).

Particle size. In the developed suppositories, the particle sizes under the dispersion technique used in laboratory conditions were medium-dispersed, while this indicator for comparative preparations (Flagil, Pimafucin) for industrial production was finely dispersed. The basis influences more significantly the particle size of metronidazole: in hydrophilic suppositories, the degree of dispersion of metronidazole is much higher than that of diphilic. The variation in particle diameter is less in hydrophilic substrates.

The use of the Colloid Mill module (Labor Pilot homogenizer, IKA) as part of the production line made it possible to obtain suppositories with a particle size of not more than 1 μm.

Thus, the dispersion of the resulting drug is comparable with the comparison drugs.

Expedited storage. The compatibility of the components of the developed suppository formulations was investigated during accelerated storage. Suppositories were placed in a thermostat at a temperature of 30 ° C. After 6 months of storage, the following parameters were evaluated: appearance, uniformity at the cut, pH, time of dissolution or complete deformation. Assessment of physico-chemical parameters was carried out according to the article "General methods of analysis" of the American Pharmacopoeia, 26th ed. (USP-26) and the Suppositories article of the State Pharmacopoeia, ed. XI (GF-XI).

In accordance with the invention, a method for the treatment of infectious gynecological diseases, including vaginitis, vulvitis, vulvovaginitis caused by anaerobic bacteria, fungi and protozoa, is described.

As can be seen from table 10 of the "Examples" section, the antibacterial activity against the anaerobic flora of the combinations proposed in accordance with the invention exceeds that of the analogue, being active in dilution up to 1: 10000 (25 μg / ml). Natamycin increases the antibacterial effectiveness of metronidazole.

Thus, the drug has a pronounced antiprotozoal, microbicidal and antifungal effect in the experiment on representatives of the main types of infectious vaginitis pathogens, the most common among the population: Trichomonas, Bacteroides, Fusobacterium, Candida. The effectiveness of the developed drug is not inferior to monopreparations of metronidazole (Flagil) and natamycin (Pimafucin), and in relation to trichomonads it is superior.

According to another aspect of the present invention, a method for producing suppositories is described, which consists in fusing the components of a base at 30-75 ° C, grinding the solid components with a molten base, packaging and cooling.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1. The dependence of shear stress on the shear rate of the developed hydrophilic composition (Example 1) and Pimafucin.

Figure 2. The dependence of shear stress on the shear rate of the developed diphilic composition (Example 3) and Flagil.

The following are examples of embodiments of the invention, which are provided for illustration and do not limit the scope of the invention.

MODES FOR CARRYING OUT THE INVENTION

Example No. 1.

Suppositories for intravaginal administration containing metronidazole 26%, natamycin 4.75%, lidocaine hydrochloride 0.45%, taurine 0.5%, propylene glycol 0.85%, PEG 1500 - 0.9%, emulsifier T-2 - 4, 5%, tween-80 - 1.5% and solid fat type A - 50.45%.

The manufacturing method includes the following main steps.

1. Weighing components, preparation of equipment.

2. Preparation of a solid fat melt with T-2 and a PEG melt with tween 80 and propylene glycol.

3. Mixing solids with melts, simultaneous mixing and grinding in a homogenizer.

4. Dosing in foil blisters.

5. Cooling.

Example No. 2.

Suppositories for intravaginal administration containing metronidazole 26.5%, natamycin 5.75%, lidocaine hydrochloride 0.01%, taurine 1%, propylene glycol 4.73%, PEG 1500 - 1%, emulsifier T-2 - 5%, tween -80 - 2.0% and solid fat type A - 50.45%. The cooking steps correspond to example 1.

Example No. 3.

Suppositories for intravaginal administration containing metronidazole 26.0%, natamycin 4.75%, lidocaine hydrochloride 0.45%, taurine 0.5%, propylene glycol 0.85%, PEG 1500 - 65.25%.

The manufacturing method includes the following steps.

1. Weighing components, preparation of equipment.

2. Preparation of a PEG melt with propylene glycol.

3. Mixing solids with melts, simultaneous mixing and grinding in a homogenizer.

4. Dosing in aluminum foil blisters.

5. Cooling.

Example No. 4.

Suppositories for intravaginal administration containing metronidazole 26.5%, natamycin 5.75%, lidocaine hydrochloride 0.5%, taurine 0.02%, propylene glycol 0.01%, polyethylene glycol 1500 - 65.25%. The cooking steps correspond to example 3.

Example No. 5.

Suppositories for intravaginal administration, containing tinidazole 0.5 g, amphotericin 0.1 g, benzocaine 0.02 g, arginine 0.1 g, hydrophilic base 1.48 g.

The manufacturing method includes the following main steps.

1. Weighing components, preparation of equipment.

2. Melt preparation of PEG 1500 with propylene glycol at 50-60 ° C.

3. Mixing solids with the molten base, simultaneous mixing and grinding in a homogenizer for 10-15 minutes.

4. Dosing in aluminum foil blisters.

5. Cooling.

The composition is stable, particle size within 10 microns, average dissolution time 22 minutes.

Example No. 6.

Suppositories for intravaginal administration, containing orniadzole 0.5 g, nystatin 0.015 g, bupivacaine 0.01 g, lysine 0.05 g, diphilic base 1.67 g.

The manufacturing method includes the following main steps.

1. Weighing components, preparation of equipment.

2. Preparation of a solid fat melt with T-2 emulsifier and a PEG melt with tween 80 and propylene glycol.

3. Mixing solids with the molten base, simultaneous mixing, partial dissolution and grinding in a homogenizer for 10-15 minutes.

4. Dosing in aluminum foil blisters.

5. Cooling.

The composition is stable, particle size within 20 microns, the average value of the period of complete deformation of 8 minutes.

Example No. 7.

Suppositories for intravaginal administration containing metronidazole 26%, natamycin 4.75%, lidocaine hydrochloride 0.45%, taurine 0.5%, propylene glycol 0.85%, PEG 1500 - 0.9%, emulsifier T-2 - 4, 5%, tween-80 - 1.5% and solid fat type A - 50.4%, ascorbate buffer 0.2%.

The manufacturing method includes the following main steps.

1. Weighing components, preparation of equipment.

2. Preparation of a solid fat melt with T-2 emulsifier and a PEG melt with tween 80 and propylene glycol.

3. Mixing solids with melts, simultaneous mixing and grinding in a homogenizer.

4. Dosing in aluminum foil blisters.

5. Cooling.

Example No. 8.

Suppositories for intravaginal administration containing metronidazole 26%, natamycin 4.75%, lidocaine hydrochloride 0.45%, taurine 0.5%, propylene glycol 0.85%, PEG 1500 - 0.9%, emulsifier T-2 - 4, 5%, tween-80 - 1.5% and solid fat type A - 50.4%, oxalate buffer 0.2%, EDTA 0.1%.

The manufacturing method includes the following main steps.

1. Weighing components, preparation of equipment.

2. Preparation of a solid fat melt with T-2 and a PEG melt with tween 80 and propylene glycol.

3. Mixing solids with melts, simultaneous mixing and grinding in a homogenizer.

4. Dosing in aluminum foil blisters.

5. Cooling.

Example No. 9.

Suppositories for intravaginal administration, containing metronidazole 26.0%, natamycin 4.75%, lidocaine hydrochloride 0.45%, taurine 0.5%, propylene glycol 0.85%, PEG 1500 - 65.1%, ascorbate buffer 0.2 %

The manufacturing method includes the following steps.

1. Weighing components, preparation of equipment.

2. Preparation of a PEG melt with propylene glycol.

3. Mixing solids with melts, simultaneous mixing and grinding in a homogenizer.

4. Dosing in aluminum foil blisters.

5. Cooling.

Example No. 10.

Suppositories for intravaginal administration containing metronidazole 26.0%, natamycin 4.75%, lidocaine hydrochloride 0.45%, taurine 0.5%, propylene glycol 0.85%, PEG 1500 - 65.1%, EDTA 0.2% .

The manufacturing method includes the following steps.

1. Weighing components, preparation of equipment.

2. Preparation of a PEG melt with propylene glycol.

3. Mixing solids with melts, simultaneous mixing and grinding in a homogenizer.

4. Dosing in aluminum foil blisters.

5. Cooling.

EXAMPLES OF RESEARCH OF PROPERTIES OF SUPPOSITORIES

Monronodications of metronidazole and natamycin were used as reference preparations: Flagil manufactured by Laboratories Aventis (state registration number P No. 014269 / 01-2002) and Pimafucin (state registration number P No. 013552 / 01-2002) manufactured Yamanouchi, as well as monopreparations of metronidazole and natamycin in our chosen basics (Table 1).

Example No. 11. Disintegration in the aquatic environment

The time and nature of dissolution (formation of a homogeneous dispersion in water) of the developed diphilic and hydrophilic compositions in the aquatic environment was carried out according to the requirements of the article “Determining the Disintegration of Dosage Forms” GF-XI, issue 2, p. 158, using the “Swing Basket” device manufactured by Ervek and isotonic sodium chloride solution as a dispersion medium.

table 2
The dissolution time (dispersion formation) of metronidazole and natamycin preparations and the nature of the systems formed Structure* dissolution time, min nature of the disperse system Diphilic 47 ± 3 emulsion Diphilic without natamycin 35 ± 1 emulsion Diphilic without metronidazole 45 ± 2 coarse emulsion Hydrophilic 25 ± 3 emulsion Hydrophilic without natamycin 20 ± 2 solution Hydrophilic without metronidazole 30 ± 2 solution Pimafucin 43 ± 2 emulsion Flagil insoluble -

* The name of the composition corresponds to the name in table 1

As can be seen from table 2, the average dissolution time of the diphilic composition, or rather the formation of a homogeneous dispersion, according to the results of three experiments was 45 minutes. The dissolution time of the hydrophilic composition was 25 minutes in three consecutive experiments. Metronidazole improves the disintegration and solubility of the form in the aquatic environment. Flagyl having a lipophilic base does not form an aqueous dispersion in this test. Pimafucin forms a dispersion within 40-45 minutes. These indicators differ for the better from indicators for comparison drugs. The dissolution time of the diphilic composition and the hydrophilic composition meets the requirements of GF-X1, therefore, the compositions provide an effective distribution over the mucous membrane.

Example No. 12. (PH measurement of formulations)

The pH value of the developed formulations (combined and mono-) was compared with the pH of monopreparations: Pimafucin (natamycin only, diphilic base) and Flagil (metronidazole only, lipophilic base). Sample preparation of diphilic suppositories was carried out in accordance with GF-XI for ointments (p. 114), dispersing the suppository with glass beads in water in a ratio of 1:10 to obtain an emulsion, inclusions were dispersed by ultrasound in an Ultrasonic installation, Germany.

Instrument setup and measurements were carried out under thermostated conditions at a temperature of 20 ° С. The pH was determined by the potentiometric method (GF-XI, pp. 113-114) using an automatic pH meter of the RN-150 brand (Russia) with an accuracy of 0.01.

Table 3
PH values of metronidazole and natamycin Structure* PH average Diphilic 6.5 ± 0.2 Diphilic without natamycin 7.0 ± 0.1 Diphilic without metronidazole 6.9 ± 0.1 Hydrophilic 7.5 ± 0.2 Hydrophilic without natamycin 7.2 ± 0.1 Hydrophilic without metronidazole 7.5 ± 0 Pimafucin 5.9 ± 0.1 Flagil 6.9 ± 0.2

* The name of the composition corresponds to the name in table 1

Example No. 13. (Melting point)

The melting temperature (T _pl , ° С exp.) Was determined on a device with a capillary equipped with a Ubellode thermometer according to the method of GF-XI, issue 1, p. 18. Reliable considered indicators with a spread of not more than 1 ° C in three consecutive measurements.

The melting temperature (T _PL , ° C theory.) Of the compositions was previously set according to the formula:

where T _PL - the melting temperature of the mixture;

T ₁ , T ₂ and T ₃ - the melting temperature of the individual components;

Q ₁ , Q ₂ and Q ₃ - their content in%;

The results of theoretical calculations and practical measurements are presented in table 4.

Table 4
Melting points of formulations with metronidazole and natamycin Structure* _Mp , ° C theory. T _pl , ° C exp. Diphilic 35.0 34,4 Diphilic without natamycin 35.0 34.5 Diphilic without metronidazole 35.0 33.5 Hydrophilic 37,2 37.0 Hydrophilic without natamycin 37,2 37.0 Hydrophilic without metronidazole 37,2 36.8 Pimafucin 37,2 37.1 Flagil 34.6 33.6 * The name of the composition corresponds to the name in Table 1

Example No. 14. (Consistency of formulations developed)

Structural and mechanical properties that characterize the flow, shape retention, and internal friction of the suppository base at body temperature were evaluated by values at certain intervals. The preparations were melted in a water bath at 37 ° C, the melt was immediately transferred to the cylinder of the Reotest-2 rotational viscometer (Germany) with a cylinder-to-cylinder sensing system T ₁ -B ₃ . The cylinder was in an aquarium with continuous heating of water to 37 ± 1 ° C. The dynamic viscosity was measured at various speeds (range 4-2048 s ^-1 ) of shear, first in the direction of increase, then in the direction of decreasing speed values. Based on the obtained series of the dependence of viscosity on shear rate, shear stress values were found by the formula:

τ = γ

where τ is the shear stress, sN / m (dyne / cm);

γ is the shear rate, s ^-1 ;

η - dynamic viscosity, cP · s.

Plots of the dependence τ (γ) were constructed, where the presence of hysteresis loops and passage through the rheological optimum was noted [A. Arkusha Studies of the structural and mechanical properties of ointments in order to determine the optimal consistency. Abstract of dissertation for the degree of Cand. farm. sciences. - Kharkov, 1982. - 55 p.].

The graphs τ (log ₂ γ) of the developed composition according to example 3 (hydrophilic composition), as well as Pimafucin (Fig. 1) have a weakly expressed hysteresis loop, which is due to the predominance in the composition of surfactants with a relatively large molecular weight (polyethylene glycol in the developed composition, cetyl alcohol in Pimafucin), as part of the base, and indicates the possibility of changing the structure of the melt at constant pressure. The graphs of both developed compounds pass through the rheological optimum, which means that during the distribution of the melt during the mucous membrane during natural friction (equivalent to ^-1 ).

The absence of significant loops in the composition graphs indicates the preservation of the structure with increasing mechanical stress or the rapid recovery with a decrease in the strength of the mechanical stress. Compositions containing mainly fat as a base (Example 1, diphilic composition and Flagyl) (FIG. 2) have maximum viscosity and structure stability of the melts. At the same time, the passage of their graphs τ (log2γ) through the optimum of consistency indicates the absence of discomfort and the difficulty of distribution over the mucous membrane. The presence of a pronounced dispersed phase (more than 25%) in the developed formulations relative to comparison preparations increases the value of dynamic viscosity relative to comparison preparations, which is expressed by a steeper graph line.

Example No. 15. (Determination of particle sizes of the suspension phase)

The particle size of the suspension phase (metronidazole, natamycin) was determined using a microscope with a counting eyepiece. Suppositories were melted at 40-50 ° C, the melt was transferred to heated laboratory glass, covered with a coverslip. The drug was examined using a LOMO microscope at a magnification of x150, and a MOB (Ocular Screw Micrometer) micrometer counting grid was installed on the lens. The particle size of the dispersed phase was determined according to the method of GF-XI, described in the article "Ointment", issue 2, p.146.

Table 6
Linear particle sizes of natamycin and metronidazole preparations in the field of view Structure* D _min , μm ** D _max , μm ** D _average , microns ** Diphilic less than 0.1 10.0 no more than 5,0 Diphilic without natamycin less than 0.1 15.0 no more than 10.0 Diphilic without metronidazole less than 0.1 10.0 no more than 5,0 Hydrophilic 0.2 10.0 5,0 Hydrophilic without natamycin 0.1 5.00 2.5 Hydrophilic without metronidazole 0.2 8.0 4.0 Pimafucin less than 0.1 0.15 no more than 0.2 Flagil less than 0.1 0.50 no more than 1,0 * The name of the composition corresponds to the name in table 1 ** D _min - the minimum particle size, D _max - the maximum particle size, D _avg is the arithmetic mean of the particle size.

According to Table 6, the particle size distribution obeyed the Gauss curve; no foreign inclusions were noted. For all developed compositions, a fraction with an arithmetic mean particle size of 5-7 microns, a dispersion of about 20%, and a maximum size of 15 microns prevail. The particle sizes of all drugs fit into the requirements for suspension-type drugs for external use (from 0.1 to 100 microns).

Example No. 16 (study of the specific activity of the drug)

The kinetics of release was evaluated in the Dissolution test, the Rotating Basket instrument (Erweka DT-6) was used, the experiment was carried out in accordance with the requirements of GF-XI, p. 159. An isotonic sodium chloride solution was used as a dialysis medium. Considering the prolonged nature of the release from suppositories, 5 ml samples were taken every 30 minutes, the volume was replenished with dialysis medium.

The samples were tested for their ability to suppress the growth of standard strains of microorganisms: anaerobes: Fusobacterium necroforum, Bacteroides nodosus, fungi: Candida utilis (dilution method with medium) [Krasilnikov A.P. Handbook of antiseptics. - Minsk: Higher School, 1995. - 367 p.], As well as Trichomonas.

The study of the antitrichomonas activity of metronidazole was carried out in in vitro experiments. Trichomonas strains in association with bacteria, in particular with E. coli, were cultured on Pavlova's medium, transplanting such cultures every 2-3 days. [Experimental study of the antiprotozoal activity of pharmacological substances (guidelines) / Corr. RAMS T.A. Guskova]. Trichomonas vaginalis isolated from the lesion, reseeding from Pavlova's medium (co-growth with E. coli) into a liquid nutrient medium (MPB). The activity of Trichomonas was checked (at least 80% actively moving), observing a liquid preparation at a magnification of × 1000 with a Lumex microscope. Dilutions were prepared using the decimal method, diluting 1 ml of the dialysis medium selected in the Dissolution experiment (dissolution time 60 minutes), 9 ml of sterile isotonic sodium chloride. Next, 1 ml of the resulting solution was transferred into a test tube with 9 ml of sterile isotonic sodium chloride, obtaining a dilution of 1: 100, etc. 1 ml of Trichomonas liquid culture was introduced into a test tube with 1 ml of dilution, left for 1 h, then the preparation was transferred to glass, microscopic, observing the state of flagella and membranes. For the minimum inhibitory concentration, the maximum dilution was taken, which caused the stoppage of movement of flagella and membranes of 98% of microorganisms in the field of view. Dilutions of 1% aqueous solution of the substance metronidazole were used as a standard, based on 500 mg in one suppository, which is equivalent to 1% with 100% release in 50 ml of dialysis medium.

The compositions were studied: on a diphilic basis, metronidazole + natamycin (1), natamycin (2) and metronidazole separately (3), on a hydrophilic basis, metronidazole + natamycin (4), natamycin (5) and metronidazole separately (6), Flagil preparations (metronidazole) (7) and Pimafucin (8) for comparison.

From table 8 it follows that the standard has a total trichomonostatic effect at a dilution of 1: 1000 (0.001%). All dialysate samples have a trichomonostatic effect, most pronounced for complex preparations of metronidazole + natamycin. The lipophilic base (Flagil) has the least releasing ability with respect to metronidazole - about 10% per 1 hour, hydrophilic and diphilic bases release about the same amount of metronidazole in about 1 hour - about 50%. Natamycin potentiates the trichomonacid action of metronidazole. Pimafucin has a weak trichomonacid effect at a dilution of 1: 100.

Pathogenic fungal activity was evaluated on Candida utilis strains in accordance with the GF-XI method (serial dilution method with medium). Stepwise dilutions of aqueous extracts from preparations were mixed with a nutrient medium onto which test cultures of microorganisms were seeded. The test results of serial dilutions of the drug compared with natamycin monopreparations are presented in Table 9.

Obviously, in the fungicidal efficacy of natamycin in the combinations we have proposed, it is not inferior to the analogue, showing activity in dilution to 31.25 μg / ml. Metronidazole does not affect the anti-candidiasis activity of natamycin.

Efficacy against anaerobic bacterial microflora (metronidazole) was examined on pathogenic strains of Bacteroides and Fusobacterium based on FGU VGNKI.

During storage, the compositions were tested for the absence of growth of pathogenic strains according to the GF-XI method for the absence of growth of microorganisms of the groups Pseudomonas, Enterobacteriaceae, Staphylococcus. The compositions were suspended with sterile water 1:10, kept in an ultrasonic bath for homogenization, then they were directly inoculated on special nutrient media: thioglycolic (Pseudomonas aeruginosa group), Saburo (mushrooms) and MPA (cocci). Within 1 week there was no growth of colonies of microorganisms, which indicates the preservation of microbiological purity over the expiration date.

Example No. 17 (acute and chronic toxicity)

The toxicity of the drugs, acute and chronic, was evaluated on the basis of the State Scientific Center for Antibiotics (FSUE SSCA). The experiment was conducted on rats of the pure Wistar line with daily rectal administration of 200 mg of suppository mass with simultaneous placebo control (groups of 50 and 10 animals, respectively). [Guidelines for the study of the general toxic effects of pharmacological substances. / E.V. Arzamastsev, T.A. Guskova, I.V. Berezovskaya, B.I. Lyubimov, etc. - M., 2000.]

Evaluation of the effect of the drug was carried out during the course of administration — on the general condition of the animals, immediately after the end of the administration course, and after 2 weeks — on biochemical blood parameters, the picture of peripheral blood and blood coagulability, as well as morphometric analysis and histopathological examination of the internal organs of animals. The experiments were conducted on four experimental groups of rats, 15 females and 15 males in each group.

In an acute toxicity experiment, it was found that exceeding the estimated dose of the drug by 10 times does not lead to pathological changes in the body.

No deaths or acute poisoning, biochemical parameters (total protein level, ratio of protein fractions and total serum cholesterol, glucose, urea, creatinine, or activity were recorded within 2 weeks (based on the expected duration of therapy of 6-9 days) alanine and aspartate transaminases, alkaline phosphatase) remained normal. Histological examination of the organs did not reveal abnormalities in their structure.

The study of the local irritating effect of the test drug was carried out in experiments (introduction according to the scheme prescribed for humans - 6-7 days, no more). At the end of the experiment, an examination was made of the condition of the vaginal mucosa.

No cases of irritating effects were noted, as well as modifications of the vaginal epithelium. Application of the composition to the mucous membrane of the rabbit eye (N = 5) also did not cause irritation. It was found that the developed formulations in the form of suppositories do not have an irritating effect.

Given the dose level of toxic effects, the developed suppositories of a combined composition according to the degree of danger can be classified as a low-hazard drug (degree of toxicity according to GOST 12.1.00.7.76-IY), which can be recommended as a medicine for medical use. The ratio was established experimentally and is optimal according to the results of physico-chemical, pharmacotechnological and biological studies.

Thus, the developed suppository formulations for intravaginal use have:

1) biological safety, manifested in the absence of irritating and toxic effects;

2) biocidal efficacy against specific strains of pathogens, including protozoa, fungi and anaerobic microflora;

3) the high dispersion and bioavailability of metronidazole and natamycin, comparable with the effectiveness of the monopreparations “Flagil” and “Pimafucin”;

meet the requirements of normative and technical documentation for the quality of suppositories.

Claims (12)

1. Suppositories for the treatment of infectious gynecological diseases, containing as an antibacterial and antimicrobial agent an effective amount of a compound from a number of nitroimidazoles, as an antifungal agent an effective amount of a polyene macrolide compound, and an effective amount of a local anesthetic in a diphilic or hydrophilic suppository base. 2. Suppositories according to claim 1 in the following ratio of components, wt.%:
compound from a number of nitroimidazoles 15.0-30.0 macrolide polyene compound 4.0-6.0 local anesthetic 0.01-5.0 diphilic or hydrophilic suppository base rest 3. Suppositories according to claim 2, additionally containing a reparative agent in the following ratio of components, wt.%:
compound from a number of nitroimidazoles 15.0-30.0 macrolide polyene compound 4.0-6.0 local anesthetic 0.01-5.0 reparative agent 0.01-5.0 diphilic or hydrophilic suppository base rest 4. Suppositories according to claim 3, additionally containing a buffer system in the following ratio of components, wt.%:
compound from a number of nitroimidazoles 15.0-30.0 macrolide polyene compound 4.0-6.0 local anesthetic 0.01-5.0 reparative agent 0.01-5.0 buffer system 0.01-1.0 diphilic or hydrophilic suppository base rest 5. Suppositories according to claim 3, additionally containing an antioxidant in the following ratio of components, wt.%:
compound from a number of nitroimidazoles 15.0-30.0 macrolide polyene compound 4.0-6.0 local anesthetic 0.01-5.0 reparative agent 0.01-5.0 antioxidant 0.01-2.0 diphilic or hydrophilic suppository base rest 6. Suppositories according to any one of claims 1 to 5, containing metronidazole, tinidazole, secnidazole or ornidazole as a compound from a series of nitroimidazoles; as the macrolide polyene compound, natamycin, nystatin, amphotericin or rifamycin; as a local anesthetic, lidocaine hydrochloride, xylocaine, bupivacaine, novocaine or benzocaine; as a reparative agent, amino acids selected from the group consisting of taurine, arginine, tryptophan, lysine, proline or histidine. 7. Suppositories according to any one of claims 1 to 5, containing, as a diphilic suppository base, a mixture of solid fat, T-2 emulsifier, Tween-80, PEG-500 and propylene glycol at a base content, wt.%:
Solid fat 55.0-80.0 Emulsifier T-2 4,5-6,0 Twin 80 1.5-3.0 PEG-1500 0.1-5.0 Propylene glycol 0.1-10.0 8. Suppositories according to any one of claims 1 to 5, containing as a hydrophilic suppository base a mixture of PEG-1500 and propylene glycol at a base content, wt.%:
PEG-1500 60.0-85.0 Propylene glycol 0.1-20.0 9. The suppositories according to claim 4, in which under the buffer systems added to the composition for adjusting the pH, mean ascorbate, carbonate, phosphate, citrate, oxalate buffers. 10. Suppositories according to claim 5, in which the antioxidants are selected from the group comprising ascorbic acid, ascorbyl palmitate, hypophosphoric acid, sodium ascorbate, vitamins C and E, sodium bisulfite, EDTA, citric acid and other compounds known to those skilled in the art. antioxidant properties in polymer diphilic or hydrophilic bases. 11. A method of treating infectious gynecological diseases, including the introduction of suppositories according to claims 1 to 10. 12. The method according to claim 10, where infectious gynecological diseases include vaginitis, vulvitis, vulvovaginitis caused by anaerobic bacteria, fungi and protozoa.

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