RU2367455C2 - Antibacterial and necrolytic external and local pharmaceutical composition - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: external and local pharmaceutical composition possesses antibacterial and necrolytic action, contains an active complex of bacteriolytic and proteolytic enzymes - Lisoamidase, a base and at least one desired additive chosen from: a substance disarranging the membrane structure of gram-negative microorganisms, an antibiotic, an anesthetic, a reparative process stimulator or physiologically acceptable mixture thereof. The components are taken in certain mass ratio.

EFFECT: extended application, enhanced therapeutic activity and ease of use, lower consumption of the active substance, ensured anaesthetising effect, additional stimulation of reparative processes, improved microcirculation, reduced secondary tissue necrosis surrounding a wound caused by excess free radical oxidation.

7 cl, 14 ex

Description

The invention relates to medicine and veterinary medicine, namely to pharmacological preparations used in the treatment of purulent wounds and burns, purulent-inflammatory skin diseases, etc., to accelerate and improve healing conditions.

Bacteriolytic enzyme complexes are known that lyse gram-positive microorganisms resistant to antibiotics (patents RU 1549227, C12N 9/00, 1984.10.04; RU 2193063, C12N 9/00, 2000.11.29).

The complex of bacteriolytic enzymes (patent RU 1549227) used in medicine under the name lysoamidase, and veterinary medicine - lysomast (patent RU 2064299, A61K 35/74, 1993.12.13), includes three bacteriolytic enzymes immobilized on the polysaccharide, which increases their stability. The polysaccharide provides long-term (for several years) preservation of the high activity of the enzymes included in it, as well as the immunostimulating properties of lysoamidase.

On the basis of this complex, a pharmacological preparation with antibacterial and necrolytic action has been developed, produced in the form of lyophilized powder, packaged in 10 ml vials of 50 PE, 100 PE with hermetically sealed rubber stoppers. In this case, the delivery package includes a solvent in ampoules of 5 ml per 1 ampoule per 50 PE (pharmacopoeial articles FS 42-3329-96 and FS 42-3332-96). The drug is used in liquid form in a phosphate buffer solution with a concentration of 5 PE (50 LE) in 1 ml; 10 PE (100 LE) in 1 ml; 25 PE (250 LE) in 1 ml.

Bacteriolytic enzymes included in the active complex of the drug destroy cells of gram-positive pathogenic microorganisms, such as staphylococci and streptococci, including those with multiple antibiotic resistance: radio-resistant micrococci, meningococci, gonococci, gram-positive bacilli. Due to the presence of proteases and enzymes in the complex, which also have proteolytic activity, the drug cleans wounds from necrotic masses well. Fragments of destroyed cells have an immunostimulating effect.

Clinical studies of the drug showed that lysoamidase in the treatment of purulent wounds of soft tissues accelerates the course of the wound process compared to other traditional and non-traditional means. The microflora content decreases faster, the cytogram normalizes faster, necrotic mass rejection is faster, granulation and even tissue epithelization are faster. The terms of treatment of patients with purulent wounds of soft tissues were reduced in many cases by 2 times. Lizoamidase stimulates reparative processes, exerting, in addition to necrolytic and bacteriolytic actions, an effect on the course of the 2nd phase of inflammation, not only due to the reduction of the 1st phase, which is typical for all enzyme preparations used for treatment in such cases, but also for due to the stimulating effect on the cells of granulation tissue, acting directly on the cells of the proliferate.

However, these complexes of bacterio- and proteolytic enzymes have little or no effect on gram-negative microflora, since the surface structures of gram-negative bacteria are a barrier to the lytic action of lysoamidase, and therefore the lysis process either lengthens or does not occur at all.

The use of the known drug in the form of liquid solutions has several disadvantages, namely: the drug is poorly fixed and held on the wound surface, which leads to increased consumption of the active substance during treatment; requires preliminary dilution in a buffer solution and special measures to prevent rapid drying, which is not convenient during the treatment process. All this reduces the patient's commitment (motivation) to treatment.

A known preparation for the lysis of cells of gram-positive and gram-negative microorganisms containing a complex of bacterio- and proteolytic enzymes - lysoamidase with polymyxin B (patent RU 2139348) and a buffer solution in the following ratio of components, μg / ml (%): lysoamidase 1000-3000 (0.1 -0.3), polymyxin B 20-50 (0.002-0.005), buffer solution - the rest.

In the well-known drug, together with lysoamidase, the target additive, polymyxin B, was used. This antibiotic disrupts the cell membrane of gram-negative microorganisms, which leads to the death of their cells. Instead of divalent cations, this substance binds electrostatically to the negatively charged membranes of many gram-negative bacteria. The hydrophobic fatty acid chain of polymyxin enters the hydrophobic region of the membrane, dramatically increasing its permeability, which causes a violation of the integrity of the membrane.

The disadvantage of this drug is that with prolonged use of polymyxin together with a complex of bacterio- and proteolytic enzymes, the resistance of microorganisms to the action of this antibiotic may occur.

The use of the known drug in the form of liquid solutions has several disadvantages, namely: the drug is poorly fixed and held on the wound surface, which leads to increased consumption of the active substance during treatment; requires preliminary dilution in a buffer solution and special measures to prevent rapid drying, which is inconvenient in the treatment process. All this reduces the patient's commitment to treatment.

The disadvantages of this drug is that it does not have a local anesthetic effect, does not improve microcirculation during the inflammatory process, does not have an antioxidant effect.

The objective of the invention is the creation of a pharmacological preparation based on a complex of bacteriolytic and proteolytic enzymes in the form of a soft dosage form.

The technical result that can be obtained using the proposed invention: expanding the scope, increasing therapeutic activity; improved usability; reduced consumption of active substance; obtaining an anesthetic effect; additional stimulation of reparative processes; improved microcirculation, reduced secondary tissue necrosis in the circumference of the wound associated with excessive free radical oxidation.

To solve this problem, the present invention provides a topical pharmacological preparation in the form of a soft dosage form, consisting of an active complex of bacteriolytic and proteolytic enzymes, for example, lysoamidase (hereinafter “active complex”), as well as compatible with the active complex of target additives and a base that provides soft dosage form of the drug, in the following ratio of components (wt.%):

active complex 0,100-5,000 target additive 0.002-12,000 the basis rest

As a target additive, a substance that violates the structure of the membranes of gram-negative microorganisms can be used. In this case, the drug contains these components in their following ratio (wt.%):

active complex
structural breaker 0,100-5,0 membranes of gram-negative microorganisms 0.002-2.0 the basis rest

As a substance that violates the structure of the membranes of gram-negative microorganisms, for example, Trilon B or ethyldiaminetetraacetic acid or a mixture thereof can be used.

The proposed pharmacological preparation containing the active complex, the target additive in the form of a substance that violates the structure of gram-negative microorganisms, and the base, in addition, additionally contains a local anesthetic substance as the target additive in the following ratio of components (wt.%):

active complex
structural breaker 0.1-5.0 membranes of gram-negative microorganisms, 0.002-2.0 local anesthetic 0.5-5.0 the basis rest

A local anesthetic is, for example, quinoxycaine, trimecaine, pyromecaine, lidocaine, anestezin, dibucaine, or a mixture thereof.

The proposed pharmacological preparation containing an active complex, a base and a target additive in the form of a substance that violates the structure of the membranes of gram-negative microorganisms, additionally contains a stimulator of reparative processes as a target additive in the following ratio of components (wt.%):

active complex
structural breaker 0.1-5.0 membranes of gram-negative microorganisms 0.002-2.0 reparative stimulant 0.5-5.0 the basis rest

The proposed pharmacological preparation containing an active complex, base and target additives in the form of a substance that violates the structure of the membranes of gram-negative microorganisms, and a local anesthetic substance, additionally contains as a target additive a stimulant of reparative processes in the following ratio of components (wt.%):

active complex
structural breaker 0.1-5.0 membranes of gram-negative microorganisms 0.002-2.0 local anesthetic 0.5-5.0 reparative stimulant 0.5-5.0 the basis rest

In this case, the stimulator of reparative processes can be, for example, methyluracil, acetamine, etaden, calcium pantothenate, solcoseryl, trekrezan, or a mixture thereof.

In accordance with the invention, soft dosage forms based on a complex of bacteriolytic and proteolytic enzymes, such as lysoamidase, are ointment-like soft dosage forms: ointments, gels, creams, and also formed soft dosage forms - suppositories and gelatin capsules for rectal administration.

As a hydrophilic base in a preparation containing an active complex of bacterio- and proteolytic enzymes, a physiologically acceptable mixture of hyperosmolar substances is proposed.

The following bases are proposed in the form of mixtures of hyperosmolar substances for ointment-like soft dosage forms - ointments, creams, gels, and for the formed soft dosage forms - gelatin or other capsules for rectal administration:

Base No. 1 (wt.%):

proxanol-268 or polyethylene oxide-1500 16.0-24.0 1,2-propylene glycol 46.0-50.0 PEO-400 polyethylene oxide rest

Base No. 2 (wt.%):

proxanol-268 or polyethylene oxide-1500 16.0-40.0 1,2-propylene glycol 20.0-50.0 PEO-400 polyethylene oxide 15.0-30.0 phosphate buffered saline or purified water rest

Base No. 3 (wt.%):

proxanol-268 or polyethylene oxide-1500 16.0-40.0 1,2-propylene glycol 20.0-50.0 PEO-400 polyethylene oxide 15.0-30.0 dimexide 0.5-3.0 phosphate buffered saline or purified water rest

Base No. 4 (wt.%):

polyethylene oxide-1500 or proxanol-268 16.0-24.0 PEO-400 polyethylene oxide rest

Base No. 5 (wt.%):

polyethylene oxide-1500 or proxanol-268 16.0-40.0 PEO-400 polyethylene oxide 30.0-80.0 phosphate buffered saline or purified water rest

Base No. 6 (wt.%):

polyethylene oxide-1500 or proxanol-268 16.0-40.0 PEO-400 polyethylene oxide 30.0-80.0 dimexide 0.5-3.0 phosphate buffered saline or purified water rest

Base No. 7 (wt.%):

petroleum jelly 15.0-25.0 glycerol 10.0-20.0 1,2-propylene glycol 10.0-20.0 emulsifier No. 1 5.0-15.0 phosphate buffer 10.0-15.0 purified water rest

Base No. 8 (wt.%):

polyethylene oxide PEO-1500 or proxanol-268 hyaluronic kilota 30.0-80.0 or sodium hyaluronate 0.01-3.0 phosphate buffered saline or purified water rest

Base No. 9 (wt.%):

polyethylene oxide PEO-1500 or proxanol-268 20.0-50.0 PEO-400 polyethylene oxide 30.0-10.0 hyaluronic acid or sodium hyaluronate 0.01-3.0 phosphate buffered saline or purified water rest

Base No. 10 (wt.%):

polyethylene oxide PEO-1500 or proxanol-268 20.0-35.0 PEO-400 polyethylene oxide 5.0-30.0 1,2-propylene glycol 10,0-40,0 hyaluronic acid or sodium hyaluronate 0.01-3.0 phosphate buffered saline or purified water rest

In addition, the proposed framework in the form of mixtures of hyperosmolar substances for the formed soft dosage forms - suppositories:

Base No. 11 (wt.%):

proxanol-268 or polyethylene oxide-1500 70.0-90.0 1,2-propylene glycol 5.0-20.0 PEO-400 polyethylene oxide rest

Base No. 12 (wt.%):

proxanol-268 or polyethylene oxide-1500 70.0-90.0 1,2-propylene glycol 5.0-10.0 PEO-400 polyethylene oxide 5.0-10.0 phosphate buffered saline or purified water rest

Base No. 13 (wt.%):

proxanol-268 or polyethylene oxide-1500 70.0-90.0 1,2-propylene glycol 5.0-10.0 PEO-400 polyethylene oxide 4.0-8.0 dimexide 0.5-3.0 phosphate buffered saline or purified water rest

Base No. 14 (wt.%):

proxanol-268 or polyethylene oxide-1500 70.0-90.0 PEO-400 polyethylene oxide rest

Base No. 15, (wt.%):

proxanol-268 or polyethylene oxide-1500 70.0-90.0 PEO-400 polyethylene oxide 5.0-25.0 phosphate buffered saline or purified water rest

Base No. 16 (wt.%):

proxanol-268 or polyethylene oxide-1500 70.0-90.0 PEO-400 polyethylene oxide 5.0-25.0 dimexide 0.5-3.0 phosphate buffered saline or purified water rest

As a substance that violates the structure of the membranes of gram-negative microorganisms, the proposed solution used ethylenediaminetetraacetic acid (EDTA), its disodium salt (Trilon B), or a mixture thereof.

EDTA reduces the resistance of bacteria to antibiotics and other antibacterial substances, especially for which calcium and magnesium ions are antagonists of their antimicrobial effects, such as polymyxins and aminoglycosides. Therefore, EDTA can be used in conjunction with polymyxins in the proposed drug, for example polymyxin B, to reduce the resistance of microorganisms to them. With prolonged contact of the EDTA solution with various types of microorganisms in the cells of the latter, the destruction of nucleic acids occurs, which leads to their lysis. In addition to affecting the cell membrane and bacterial metabolism, EDTA induces the loss of plasmids in staphylococcus transductant strains, and also inhibits the genetic transfer of R-plasmids in staphylococci, which inhibits transduction of resistance, for example, to erythromycin 200 times.

Trilon B has a pronounced damaging effect on the microbial cell, since it is a chelating substance that binds divalent metal ions, which causes the membrane to lose the lipopolysaccharide complex. As a result of this, the cell wall becomes fragile, its permeability to various substances, including antimicrobial action, increases. Trilon B exhibits a bacteriostatic effect in relation to strains of gram-negative and gram-positive pyogenic bacteria: staphylococcus and bacillus of blue-green pus at a concentration of 0.075%; Protein and Escherichia coli 0.15% ("Theory and practice of local treatment of purulent wounds." / Ed. by Prof. B. M. Dotsenko. Kiev, "Health", 1995). Trilon B has a preservative effect for drugs.

In the proposed preparation, Trilon B is used with a complex of bacteriolytic and proteolytic enzymes for the treatment of wounds with polyvalent microflora, which includes gram-positive and gram-negative microorganisms.

Since the mechanisms of action of polymyxin B, EDTA and trilon B on gram-negative microorganisms are different, it is advisable to use these substances together in one drug to enhance the lysis of gram-negative microorganisms, for example, in the form of complexes: polymyxin B (0.001-0.005%) and EDTA (0.001- 0.02%); EDTA (0.001-0.02%) and Trilon B (0.002-0.08%); polymyxin B (0.001-0.005%), EDTA (0.001-0.02%) and Trilon B (0.002-0.08%).

The combined use of a complex of bacteriolytic and proteolytic enzymes with a substance that violates the membrane structure of microorganisms, in the form of a monocomponent or as a mixture of components, significantly enhances the antibacterial effect of the proposed drug.

Disruption of microcirculation is one of the main factors in the pathogenesis of the inflammatory process. It includes not only blood flow through arterioles, capillaries and venules, but also transcapillary exchange, which provides tissue homeostasis necessary for life. Microcyclic disorders are divided into intravascular changes, vascular disorders and extravascular changes. The therapeutic range of drugs used to improve microcirculation in wound tissues can be largely realized due to their antihistamine, anti-inflammatory and dehydrating effects.

The use of trilon B in the proposed drug provides improved microcirculation in the tissues of the wound, since trilon B reduces the activity of histamine. In addition, the use of a hyperosmolar base compatible with a complex of bacterio- and proteolytic enzymes also improves microcirculation in wound tissues due to its dehydrating effect. In this case, the applied base, consisting of hyperosmolar substances, has an indirect anti-inflammatory effect due to the absorption of fluid and reduce swelling of wound tissues.

Secondary tissue necrosis in the circumference of a wound is partially associated with excessive free radical oxidation.

Since Trilon B is a water-soluble antioxidant, its use in the proposed drug reduces secondary tissue necrosis around the wound.

One of the main tasks of local treatment of wounds and burns is the problem of suppressing pain.

To solve this problem, in the proposed preparation, a local anesthetic substance is used as the target additive, as quinoxicaine (a quinoline derivative) with bacterial and antioxidant activity; lidocaine; trimecaine, which retains its activity in an acidic environment, which is characterized by a purulent wound; pyromecain; anestezin; dibucaine.

Lidocaine, trimecaine and pyromecaine are the most promising in the composition of the proposed drug on a hydrophilic basis.

For the proposed fat-based preparation, for example, anestezin (benzocaine) and dibucaine should be used.

To provide additional stimulation of the repair processes in the wound, the proposed drug based on a complex of bacterio- and proteolytic enzymes contains a wound healing substance: methyluracil, atsemin, etaden, calcium pantothenate, solcoseryl, trekrezan, or a mixture thereof.

Methyluracil, being a pyrimidine derivative, has anabolic and anti-catabolic activity. The substance has no local irritant effect; accelerates the processes of cell regeneration, reducing the healing time of wounds; stimulates cellular and humoral defense factors; has an anti-inflammatory effect; stimulates erythro- and leukopoiesis.

Acemine accelerates tissue regeneration and epithelization processes. It is used to treat long-term non-healing wounds and burns.

Etaden stimulates reparative processes in the epithelial and hematopoietic tissues, activating the metabolism of nucleic acids. It is indicated for the treatment of thermal and radiation injuries of the skin, rectal mucosa, nonhealing granulating wounds, etc.

Calcium pantothenate is a promising wound healing agent. It is used to treat burns and sluggishly healing wounds.

Solcoseryl is a cattle blood extract freed from proteins and lacking antigenic activity. The substance accelerates the growth of granulations and epithelization of wounds, improves metabolic processes in tissues.

Trekrezan is a triethanolammonium salt of 2-methylphenoxyacetic acid. It stimulates proliferative and reparative processes, which makes it promising for use in surgical practice to accelerate the healing of purulent necrotic injuries and postoperative wounds (MG Voronkov, MM Rasulov. “Trekrezan is the founder of a new class of adaptogens and immunomodulators (review)” Chemical and Pharmaceutical Journal, 2007, Volume 41, No. 1, pp. 3-7).

From the microbiological point of view, in connection with the presence of external polysaccharide membranes and hydrophilic capsules in gram-negative and gram-positive bacteria, which prevent the penetration of hydrophobic antibacterial substances into microbial cells, hydrophilic bases are preferable, since they serve as drug conductors into the microbial cell.

Hydrophilic bases, which can be used with a complex of bacteriolytic and proteolytic enzymes, for example, lysoamidase, in the proposed preparation are divided into two groups:

- water-soluble bases, which, as a rule, contain purified water, hydrophilic non-aqueous solvents: polyethylene oxide, for example, PEO-400, 1,2-propylene glycol, glycerin, ethyl cellosolve, dimexide;

- and water-soluble polymers: polyethylene oxide, for example PEO-1500, proxanol, for example, proxanol-268;

- water-washable bases, which in addition to water-soluble polymers and hydrophilic non-aqueous solvents contain lipophilic substances: higher fatty alcohols, petroleum jelly, liquid paraffin, lanolin, waxes (these bases are usually oil / water emulsions and require the presence of an oil / water emulsifier) .

Basics with hyperosmolar properties play an important role in local therapy of purulent wounds. They provide an outflow of purulent exudate from the wound, have an anti-inflammatory effect due to the absorption of fluid and reduce swelling of the wound tissue, and also protect the tissue from overhydration. Often they contribute to enhancing the action of antibacterial substances.

Substances with high hyperosmolar properties include polyethylene oxides - water-soluble products of the polymerization of ethylene oxide, which are physiologically indifferent compounds. This class of substances includes hydrophilic non-aqueous solvents (polyethylene oxide, for example, PEO-400; 1,2-propylene glycol; glycerin; ethyl cellosolve; dimexide) and polymers (polyethylene oxide, for example PEO-1500; proxanol-268). They are easily applied to the wound surface and evenly distributed on it, improving the contact of the ointment with the tissues and contents of the wound, mix well with wound exudate and maintain their uniformity, low toxicity and do not irritate tissues, do not violate their physiological functions.

In a purulent wound, PEOs of different molecular weights behave differently. So, PEO-1500 molecules stably maintain their primary localization: they remain in the wound cavity and actively bind the inflammatory exudate, giving it to the dressing with which the liquid evaporates. The released PEO-1500 molecules reattach water to themselves, and the process repeats. Smaller PEO-400 molecules have the ability to penetrate deep into the tissues and after 2 hours are determined in the bottom and walls of the wound. Forming complexes with antibacterial substances, PEO-400 conducts them into wound tissues, where microbes are localized.

The ability of polyethylene oxides to provide an antimicrobial drug with an interstitial effect fundamentally distinguishes this ointment base from fatty bases or solutions of antibiotics (antiseptics), the antimicrobial effect of which is limited only by the wound surface, although it is the deep microflora that determines the severity and direction of the purulent process.

The main property of PEO is that they have a pronounced dehydrating (moisture-absorbing) effect on the tissue. Due to the high ability to hydrate, these polymers actively absorb wound exudate, and with it microbial toxins, tissue breakdown products, as well as various biologically active substances - lysosomal enzymes and other inflammatory mediators, thereby interrupting its progressive course. Of fundamental importance is the fact that in a purulent wound, the dehydrating effect of PEO applies not only to the tissues that form the bottom and walls of the wound cavity, but also to microbial cells contained in the wound.

Dehydration of microbial cells leads to a significant decrease in their biological activity and, in particular, resistance to the action of certain drugs. Therefore, in the presence of PEO, the antimicrobial effect of antibiotics is increased tenfold. It is also important that the dissolution of antibiotics, sulfanilamides and other substances in PEO leads to an increase in their dispersion with an increase in therapeutic activity, which is also facilitated by a faster release of antimicrobial substances from the base.

PEO is an active conductor of drugs across the skin barrier. The important thing is that this penetrating without damage to the cells ability of PEO is enhanced in conditions of inflammation.

Hydrophilic bases on polyethylene oxides are able to dissolve in the secrets of the mucous membranes, completely release medicinal substances, do not irritate the mucous membranes, have a long shelf life, high physiological indifference, which contributes to their use in ointment-like and formed soft dosage forms.

An important biomedical requirement for ointment soft dosage forms is the ability to wet the wound surface and spread over it, due to the surface-active properties of solvents, polymers, surfactants and the base as a whole. The most pronounced surface activity are alcohols and ethyl cellosolve, which are unsuitable for inclusion in the composition of the base in high concentrations.

Of the glycols, it is preferable to use 1,2-propylene glycol and PEO-400 compared to glycerol having weak surface-active properties.

In the State Register of Medicines, 5 polyethylene oxides (PEO) with various molecular weights are registered: PEO-400, PEO-1000, PEO-1500, PEO-2000 and PEO-4000.

When mixing the PEO-400 solvent with PEO-1500 polymer, the latter acts as a thickener, which, depending on the amount in the total mass, provides the required consistency of the created dosage form. In addition, PEO-1500 is an osmotically active substance that absorbs purulent exudate due to the binding of water molecules to hydroxyl and ether groups. This basis is recommended for ointment dosage forms used in the local treatment of wounds in the first phase of the wound process.

Proxanol-268 is a block copolymer of propylene and ethylene oxides. Proxanol-268 as an osmotically active substance provides a more uniform absorption pattern and has a longer period of osmotic action. The total mass of absorbed water with proxanol-268 solutions in 24 hours significantly exceeds that in the case of PEO-1500. This is due to the higher mass of proxanol-268 (8.7 times more than PEO-1500).

The kinetics of water absorption by solvents and polymers depends on their molecular weight, concentration and molecular structure. So PEO-400 absorbs more water than 1,2-polyethylene glycol, and has a longer period of osmotic action. Sorption activity of 1,2-propylene glycol is maintained for 4-6 hours. This is due to both its lower molecular weight in comparison with PEO-400 and the lower hydrophilic-lipophilic balance (HLB), which for propylene glycol is 9.38. The value of HLB is greater, the greater the hydrophilicity of a substance. Therefore, the higher the HLB and molecular weight, the stronger and longer the osmotic effect of a substance at a given concentration will manifest itself.

The HLB values for some substances are: 1,2-propylene glycol 9.375, glycerol 11.275, ethyl cellosolve 8.3, PEO-400 12.49, PEO-1500 20.74.

1,2-Propylene glycol, PEO-400, PEO-1500, proxanol-268 can be preservatives in the developed soft dosage forms.

Dimexide - a 30% solution of dimethyl sulfoxide (DMSO) is used as an anti-inflammatory drug for external use, inactivating hydroxyl radicals and improving the course of metabolic processes in the focus of inflammation. It also has a local anesthetic, analgesic and antimicrobial effect; has moderate fibrinolytic activity. Penetrates through the skin, mucous membranes, the membrane of microbial cells (increases their sensitivity to antibiotics) and other biological membranes without damaging them, and can carry a variety of drugs deep into the wound. The negative characteristics of dimexide include the fact that it dries quickly.

Hyaluronic acid - a polysaccharide from the glucosaminoglycan family, has the ability to bind 1000 times more water than it weighs. Hyaluronic acid has the highest hygroscopicity compared to other moisturizing agents such as glycerin and sorbitol. In this case, the absorbed water is retained inside the gel and does not evaporate even when the relative humidity of the surrounding air decreases. Due to its high hydrophilicity, hyaluronic acid forms highly viscous gels with water that moisturize the skin and mucous membranes. Hyaluronic acid is non-toxic. As part of the gels, hyaluronic acid has an antimicrobial, wound healing and anti-inflammatory effect, promotes the regeneration of the epithelium, prevents the formation of granulation tissues, adhesions, scars, normalizes blood circulation, and does not cause clinical complications.

Hyaluronic acid has a positive effect on the course of local inflammatory processes, binds the microflora, as well as the products of its decay and vital functions, has a pronounced anti-inflammatory effect and a local anesthetic effect. At the same time, hyaluronic acid also provides mechanical protection for wounds, as it forms a thin film that prevents the penetration of environmental factors, pathogenic agents, into the wound. Gels based on hyaluronic acid are used to treat local and general purulent-inflammatory processes, thermal and chemical burns, trophic ulcers in chronic venous insufficiency, radiation injuries of the skin, cracks, abrasions, as well as for the treatment of long-healing wounds of various etiologies (patent RU No. 2195262 , A61K 9/06, 2002.12.27).

Sodium hyaluronate - the sodium salt of hyaluronic acid, is readily soluble in water with the formation of a viscous colorless gel with a pH of 6.0-7.5 (0.1% solution). In its activity, sodium hyaluronate is a complete analogue of hyaluronic acid, promotes tissue regeneration without scarring, and has the ability to stimulate cell migration and proliferation. Used in concentrations ranging from 0.01

up to 0.5%.

The polyethylene oxide base of ointments has a number of disadvantages, which include the risk of damage to cell membranes, excessive dehydration of tissue cells, osmotic shock leading to their death.

Studies have shown that the use of hydrophilic solvents and polymers individually adversely affects tissue cells. Moreover, penetrating, semi-penetrating and completely penetrating hydrophilic substances have a cytotoxic effect. With a combination of solvents and polymers, the situation changes dramatically. Cells remain viable with dilutions 1-3 orders of magnitude lower. Moreover, when comparing the effect on cells, it was found that instead of PEO-1500, proxanol-268 is preferable to be used in the basics. The introduction of about 5% of easily penetrating dimexide into the base causes a significant deterioration in the viability of tissue cells.

To address these shortcomings, bases have been developed that have the mildest cytotoxic effect on tissue cells. These bases include: 1) a mixture of 1,2-propylene glycol, PEO-400 and proxanol-268 in the ratios 48:32:20; 2) PEO-400 and proxanol-268 in a ratio of 8: 2; 3) PEO-400 and PEO-1500 in a ratio of 8: 2 (the basics are indicated by the degree of increase in the cytotoxic effect on tissue cells).

The ratio between 1,2-propylene glycol and PEO-400 in a hydrophilic basis is considered an important biopharmaceutical factor that not only affects the release of drugs through a semipermeable membrane and osmotic properties, but also on which the zones of growth inhibition of bacteria depend. The prevalence of a PEO-400 base in a dispersed medium (more than 50%) adversely affects the antibacterial properties of the ointment.

Bases with proxanol-268 penetrate under the scab better than bases with PEO-1500. This is explained by the better ability to wet the wound surface and spread proxanol-268 along it, which has higher surface-active properties than PEO-1500.

The results of the studies made it possible to substantiate the compositions of hydrophilic bases with a pronounced osmotic effect, but not damaging the tissue. From this perspective, perspectives are foundations consisting of proxanol-268, PEO-400 and 1,2-propylene glycol, to which water can be added (“Theory and practice of local treatment of purulent wounds.” / Ed. By Prof. B.M. Dotsenko . Kiev, "Health", 1995, S. 256-257).

The composition of the solvents allows you to adjust the melting temperature of ointment bases, as well as the rheological properties of ointments.

The above analysis of the known properties of hyperosmolar substances and their combinations for soft dosage forms made it possible to develop the above base formulations with hyperosmolar properties for the proposed pharmacological preparation containing a complex of bacteriolytic and proteolytic enzymes.

The type of dosage form of the proposed pharmacological preparation depends on the molecular weight of the hyperosmolar substances used, their ratio in the composition of the mixture, as well as the quantitative ratio of the hyperosmolar base with other components of the pharmacological preparation. Based on these quantitative ratios, the dosage form of the proposed pharmacological preparation may be a liquid, gel, cream, ointment or suppository.

The ointment form is most preferable for external use, since it is well fixed and held on the wound surface, has longer therapeutic activity, cheaper to manufacture, and is convenient to use, which enhances the patient's motivation for treatment.

Using a complex of bacteriolytic and proteolytic enzymes, a substance that violates the structure of the membranes of gram-negative microorganisms, a local anesthetic substance and a stimulant of reparative processes, as well as the above bases for soft dosage forms, compositions of hydrophilic ointments, hydrophilic creams, hydrophilic gels, which can also be used for the manufacture of gelatin or other capsules for rectal administration, suppositories.

Hydrophilic ointments are hyperosmolar, as a result of which, when used, they can absorb a significant amount of exudate.

Hydrophilic creams are prepared on the basis of a water / oil or water / oil / water emulsion stabilized with suitable emulsifiers. These also include colloidal dispersed systems consisting of water-dispersed or mixed water-glycol solvents of higher fatty alcohols or acids, stabilized by hydrophilic surfactants. Creams are biphasic or multiphase disperse systems, the dispersion medium of which at the set storage temperature, as a rule, has a Newtonian type of flow and low values of rheological parameters.

Hydrophilic gels (hydrogels) are prepared on the basis of water, a hydrophilic mixed or non-aqueous solvent (glycerin, 1,2-propylene glycol, PEO-400 polyethylene oxide) and a hydrophilic gelling agent (carbomers, cellulose derivatives, tragacanth, sodium hyaluronate, hyaluronic acid, etc. .). Gels are single, double, or multiphase disperse systems with a liquid disperse medium, the rheological properties of which are due to the presence of gelling agents in relatively low concentrations. In this dosage form, the gelling agents can additionally play the role of stabilizers of disperse systems: suspensions or emulsions.

Hydrophilic suppositories prepared on the above bases easily dissolve in the intestine, evenly distributing medicinal substances throughout the mucosa, which have both local and resorptive effects on the body.

Rectal gelatine capsules are more promising compared to suppositories from a technological, biopharmaceutical and economic point of view. The gelatin shell protects the drug from environmental factors and has advantages over suppositories, since it can encapsulate substances in the form of ointment, liniment, cream, gel, emulsion, suspension, etc.

The release of the drug substance occurs faster and easier than in suppositories, since under the influence of a weakly alkaline medium (pH 7.3-7.6) the contents of the rectum gelatin shells swell, and in this state even a slight peristalsis of the rectal wall is sufficient to rupture and releasing content. The amount of drug substance in the capsule, which has the necessary therapeutic effect, is a double dose of suppositories.

Thus, the production of rectal agents in a gelatin shell allows you to save expensive biologically active ingredients and reduce the cost of drugs. Rectal gelatin capsules with the proposed pharmacological preparation meet all the requirements for ideal suppositories and can be used in medicine for the treatment of proctologic diseases.

A complex of bacteriolytic and proteolytic enzymes, such as lysoamidase, in the proposed pharmacological preparation lyses gram-positive microorganisms, including those resistant to antibiotics, and also, due to its proteolytic activity, cleans wounds from necrotic masses, while providing an immunostimulating effect. In addition, lysoamidase stimulates reparative processes due to the stimulating effect on granulation tissue cells.

The use in the proposed preparation of a broad-spectrum substance that violates the structure of the membranes of gram-negative microorganisms ensures the lysis of gram-negative microorganisms. At the same time, as options, the substance was proposed both as a monocomponent and as a mixture of components, each of which has its own mechanism of lysis of gram-negative microorganisms, which significantly enhances the death of their cells.

The combined use of a complex of bacteriolytic and proteolytic enzymes with a substance that violates the membrane structure of microorganisms in the form of a monocomponent or as a mixture of components significantly enhances the antibacterial effect of the proposed drug. In addition, a broad-spectrum substance, Trilon B, provides improved microcirculation in wound tissues by reducing the activity of histamine, and as a water-soluble antioxidant, it reduces secondary tissue necrosis around the wound circumference.

The use of a local anesthetic substance in the proposed preparation as a monocomponent, or as a mixture of components, significantly suppresses pain in the treatment of wounds and burns. In this case, the use of a local anesthetic substance with antibacterial and antioxidant activity, for example, quinoxicain, enhances the therapeutic effect of the proposed pharmacological preparation.

To provide additional stimulation of the repair processes in the wound in the proposed pharmacological agent, a substance stimulating the reparative processes in the wound is used both as a monocomponent and as a mixture of components with a different mechanism and spectrum of action.

The use of a hydrophilic base, consisting of a mixture of polyethylene oxides and proxanol, in the proposed pharmacological preparation provides fast delivery of active substances to both microbial cells and tissue cells, which increases the therapeutic activity of the proposed drug. In addition, dehydration with a hydrophilic base of cells of gram-positive and gram-negative microorganisms reduces their resistance to the destructive effect of a complex of bacterio- and proteolytic enzymes, as well as a substance that violates the structure of cell membranes. It also increases the therapeutic activity of the proposed drug.

The use of a hyperosmolar base additionally improves microcirculation in wound tissues due to its dehydrating effect, and also has an additional anti-inflammatory effect due to the absorption of fluid and reduction of edema of wound tissues. The hyperosmolar base provides an outflow of purulent exudate from the wound and protects the tissue from overhydration. It is easily applied to the wound surface, it is well kept on it, evenly distributed on it, improving the contact of the ointment with tissues and wound contents, penetrates well under the scab, mixes well with wound exudate, while maintaining its uniformity, low toxicity and does not irritate tissue does not violate their physiological functions.

The dissolution of active substances in a polyethylene oxide base increases their dispersion and the ability to more quickly release from the base, which also increases the therapeutic activity of the proposed drug. This base is a conductor of active substances through the skin barrier, while such a penetrating without damage to cells is enhanced in conditions of inflammation. The hydrophilic base on polyethylene oxides is able to dissolve in the secrets of the mucous membranes, completely releasing medicinal substances without irritating the mucous membrane, has a long shelf life and high physiological indifference.

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

Example 1. Ointment No. 1.

100 g of ointment contains, g:

lysoamidase 1.00 Trilon B 0.02

base number 1:

proxanol-268 19.80 1,2-propylene glycol 47.51 PEO-400 polyethylene oxide 31.67

Example 2. Ointment No. 2.

100 g of ointment contains, g:

lysoamidase 2.00 Trilon B 0.01 EDTA 0.01 polymyxin B 0.002 trimecaine 2.00 etaden 4.00

base number 2:

proxanol-268 30.00 1,2-propylene glycol 30.00 PEO-400 polyethylene oxide 20.00 phosphate buffer (0.01 M), pH 8.0 11,978

Example 3. Ointment No. 3.

100 g of ointment contains, g:

lysoamidase 2.00 Trilon B 0.01 EDTA 0.01 quinoxicaine 1.00 methyluracil 3.00 atsemin 1.00

base number 3:

proxanol-268 30.00 1,2-propylene glycol 30.00 PEO-400 polyethylene oxide 20.00 dimexide 2.00 phosphate buffer (0.01 M), pH 8.0 10.98

Example 4. Ointment No. 4.

100 g of ointment contains, g:

lysoamidase 1.00 Trilon B 0.02 methyluracil 4.00

base number 4:

proxanol-268 19.00 PEO-400 polyethylene oxide 75.98

Example 5. Ointment No. 5.

100 g of ointment contains, g:

lysoamidase 1.00 EDTA 0.04 pyromecain 2.00

base number 5:

proxanol-268 20.00 PEO-400 polyethylene oxide
phosphate buffer 70.00 (0.01 M), pH 8.0 7.96

Example 6. Ointment No. 6.

100 g of ointment contains, g:

lysoamidase 4.00 Trilon B 0.08 trimecaine 3.00

base number 6:

proxanol-268 25.00 PEO-400 polyethylene oxide 50.00 dimexide 3.00 phosphate buffer (0.01 M), pH 8.0 14.92

Example 7. Cream.

100 g of cream contains, g:

lysoamidase 2.00 Trilon B 0.05

base number 7:

petroleum jelly 22.00 glycerol 17.00 1,2-propylene glycol 12.00 emulsifier No. 1 10.00 phosphate buffer (0.01 M), pH 8.0 14.00 purified water 22.95

Example 8. The gel.

100 g of gel contains, g:

lysoamidase 1.00 Trilon B 0.02

base number 8

PEO-1500 polyethylene oxide 50.00 hyaluronic acid 0.50 purified water 48.48

Example 9. The gel.

100 g of gel contains, g:

lysoamidase 1.00 Trilon B 0.04

base number 9

proxanol-268 40.00 PEO-400 polyethylene oxide 30.00 sodium hyaluronate 0.30 phosphate buffer (0.01 M), pH 8.0 28.66

Example 10. The gel.

100 g of gel contains, g:

lysoamidase 2.00 Trilon B 0.04 trimecaine 4.00 methyluracil 2.00

base number 10:

proxanol-268 40.00 PEO-400 polyethylene oxide 15.00 1,2-propylene glycol 10.00 sodium hyaluronate 0.50 purified water 26.46

Example 11. A mixture for the preparation of suppositories.

100 g of the mixture for the preparation of suppositories contains, g:

lysoamidase 1.00 Trilon B 0.04 trimecaine 4.00 etaden 4.00

base number 11:

proxanol-268 72.00 1,2 propylene glycol 15.00 polyethylene oxide-400 3.96

300 pieces were made from this mixture. rectal suppositories in the form of a “cigar” with a diameter of 10 mm, 32 mm long, and weighing 2.7 g. The solubility of the suppositories on a hydrophilic basis is no more than 1 hour at a temperature of (37 ± 1) ° C, which meets the requirements of Global Fund XI (Art. Suppositories pg. 151-153).

Example 12. A mixture for the preparation of suppositories.

100 g of the mixture for the preparation of suppositories contains, g:

lysoamidase 1.00 Trilon B 0.04 trimecaine 4.00 etaden 4.00

base number 12:

proxanol-268 73.00 1,2 propylene glycol 5.00 polyethylene oxide-400 5.00 phosphate buffer (0.01 M), pH 8.0 7.96

300 pieces were made from this mixture. rectal suppositories in the form of a “cigar” with a diameter of 10 mm, length 32 mm, weight 2.9 g. The solubility of the suppositories on a hydrophilic basis is no more than 1 hour at a temperature of (37 ± 1) ° C, which corresponds to the requirements of Global Fund XI (Art. Suppositories pg. 151-153).

Example 13. A mixture for the preparation of suppositories.

100 g of the mixture for the preparation of suppositories contains, g:

lysoamidase 2.00 Trilon B 0.04 trimecaine 4.00 etaden 4.00

base number 13:

proxanol-268 71.00 1,2 propylene glycol 4,50 polyethylene oxide-400 5.50 dimexide 2.00 phosphate buffer (0.01 M), pH 8.0 6.96

300 pieces were made from this mixture. rectal suppositories in the form of a “cigar” with a diameter of 10 mm, 32 mm long, weighing 2.5 g. The solubility of the suppositories on a hydrophilic basis is no more than 1 hour at a temperature of (37 ± 1) ° C, which corresponds to the requirements of Global Fund XI (Art. Suppositories pg. 151-153).

Example 14. Gelatin capsules.

The ointment from example 2 is filled with 500 pieces. gelatin capsules No. 00, a volume of 0.95 ml. The disintegration of the obtained capsules is less than 20 min, which meets the requirements of the Global Fund XI (St. Capsules, p. 143).

The bacteriolytic activity of polyethylene oxide-based lysoamidase under storage conditions at a temperature of + 4 ÷ + 8 ° C (in a sealed container) is reduced by no more than 10% within 1 year.

Based on the proposed solution, a number of new drugs can be created in the form of soft dosage forms, such as ointment and formulations, containing a complex of bacteriolytic and proteolytic enzymes, such as lysoamidase.

The proposed drug based on a complex of bacteriolytic and proteolytic enzymes, a base and a substance that violates the structure of the membranes of gram-negative microorganisms, lyses both gram-positive and gram-negative microorganisms, including those resistant to antibiotics; has a high therapeutic activity; improves usability; reduces the consumption of the active substance; gives anesthetic effect; additionally stimulates reparative processes in the wound; improves microcirculation; reduces secondary tissue necrosis in the circumference of the wound associated with excessive free radical oxidation. In addition, the proposed drug can be used rectally.

The search for information did not reveal identical and similar technical solutions.

The above set of features of the proposed pharmacological preparation (composition of the drug, its dosage form and quantitative ratio of components) was not previously known, which indicates the novelty of the proposed solution. The set of essential features of the claimed pharmacological preparation for external use based on a complex of bacteriolytic and proteolytic enzymes does not follow explicitly from the studied prior art, has significant differences from the considered analogues. So far, no similar solution has been proposed, although the preparation of lysoamidase has been known since 1989 (VFS 42-1916-89 for lysoamidase, introduced in September 1989).

Therefore, the applicant believes that the claimed pharmacological preparation for topical application based on a complex of bacteriolytic and proteolytic enzymes has an inventive step.

The inventive pharmacological preparation can be recommended in the treatment of purulent wounds and burns, purulent-inflammatory skin diseases, mastitis and endometritis, proctological diseases, etc.

The proposed pharmacological preparation for topical application based on a complex of bacteriolytic and proteolytic enzymes can be used for industrial serial production of ointment-like soft forms, such as ointments, creams, gels, as well as for industrial production of formulated soft dosage forms - suppositories and gelatin capsules for rectal use.

Claims (7)

1. A pharmaceutical composition for external and local use with antibacterial and necrolytic effects, characterized in that it contains an active complex of bacteriolytic and proteolytic enzymes - lysoamidase, a base and at least one target additive selected from the series: a substance that violates the structure membranes of gram-negative microorganisms, antibiotic, anesthetic, stimulant of reparative processes or their physiologically acceptable mixture, in the following ratio of components, wt.%:
lysoamidase 0.1-5.0 target additive 0.001-12.0 the basis rest 2. The pharmaceutical composition according to claim 1, characterized in that as a substance that violates the structure of the membranes of microorganisms, it contains substances selected from the range: Trilon B, ethyldiaminetetraacetic acid or a physiologically acceptable mixture thereof. 3. The pharmaceutical composition according to claim 1, characterized in that as an antibiotic it contains substances selected from the range: polymyxin, erythromycin, or a mixture thereof. 4. The pharmaceutical composition according to claim 1, characterized in that as an anesthetic it contains substances selected from the range: quinoxicaine, trimecaine, pyromecaine, lidocaine, anestezin, dibucaine or a mixture thereof. 5. The pharmaceutical composition according to claim 1, characterized in that, as a stimulant of reparative processes, it contains substances selected from the series: methyluracil, acetamine, etaden, calcium pantothenate, solcoseryl, trekrezan or a mixture thereof. 6. The pharmaceutical composition according to claim 1, characterized in that it contains a mixture of substances, the composition of which includes hydrophilic substances selected from the series: proxanol, polyethylene oxide, propylene glycol, dimexide, glycerin, hyaluronic acid, sodium hyaluronate, in various their combinations and, if necessary, in a mixture with at least one substance from the series: phosphate buffer solution, purified water, petroleum jelly, emulsifier No. 1. 7. The pharmaceutical composition according to claim 1, characterized in that it is made in the form of a soft dosage form: ointment, gel, cream or formulated soft dosage form: suppositories or capsules for rectal administration.