EA041409B1 - PHARMACEUTICAL FOR THE TREATMENT OF ARTHROLOGICAL DISEASES - Google Patents

Description

The invention relates to the field of medicine, in particular to medicines for the treatment of arthrological diseases.

State of the art

According to the World Health Organization, diseases of the musculoskeletal system are the leading factor in disability worldwide. Such diseases limit mobility and motor skills, leading to premature termination of labor activity, reducing the ability to perform social functions.

Most of the diseases of the musculoskeletal system are disorders that affect the joints. These include osteoarthritis, rheumatoid arthritis, psoriatic arthritis, gout, ankylosing spondylitis, etc.

A characteristic feature of these diseases is the destruction of cartilage and changes in the structure of bones. Most often, diseases of the joints are affected by people over 40 years old, but today diseases are increasingly common in the younger generation.

The basis for maintaining joint function is chondromodulating therapy, in which preparations based on the sodium salt of chondroitin sulfate, a mucopolysaccharide obtained from animal tissues, are widely used today. The drug effectively affects the metabolic processes in the patient's connective tissue, including cartilage, stimulating and normalizing the biosynthesis of glycosaminoglycans.

Chondroitin sulfate (hereinafter also referred to as CS) is a high molecular weight heteropolysaccharide that belongs to the group of glycosaminoglycans, linear unbranched polymers.

The structure of the cholesterol molecule determines its polyanionic properties and participation in the processes of transport of water, amino acids and lipids in the avascular areas of the cartilage. Long chains of cholesterol, which are part of the extracellular matrix, determine the most important biomechanical properties of cartilage tissue. Chondroitin sulfate is a structural modulator that is not only synthesized by the body, but also, after administration, integrates into the structures of cartilage tissue, stimulating its synthesis and inhibiting destruction. Its timely appointment and regular use provide inhibition, stabilization and prevention of the development of destructive processes in the joint. The main function of chondroitin sulfate in the body is to retain water and nutrients in the cartilage and ensure the movement of molecules through the cartilage tissue. This is the most important property, since there is no blood in the cartilage, which provides metabolic exchange. Chondroitin sulfate is also a chondroprotector. The water it stores in the thickness of the cartilage creates good cushioning and absorbs shocks, which ultimately increases the strength of the connective tissue.

Chondroitin sulfate is a natural component of articular cartilage, which plays an important role in maintaining the necessary osmotic pressure, due to which the matrix and collagen strands are stretched. This substance has anti-inflammatory activity, acting mainly on the cellular component of inflammation, stimulates the synthesis of hyaluronic acid and proteoglycans and inhibits the action of proteolytic enzymes. Chondroitin sulfate is the main part of articular cartilage. It is formed from repeating disaccharide units of glucuronic acid and N-acetyl-O-galactosamine. In the degenerative-dystrophic process, the amount of chondroitin decreases sharply, which leads to cartilage degradation [Builova T.V. An open randomized study of the efficacy and safety of Chondroxid® in the complex treatment of patients with chronic vertebrogenic lumboischialgia//BC. 2010. No. 17. S. 1678-1686].

Today, chondroitin sulfate medicines are presented on the market in injectable and non-injectable forms.

Non-injectable forms are represented by tablets and capsules.

The drug Artra is a film-coated tablet, 500 mg + 500 mg: 30, 60, 100 or 120 pcs. Active ingredients: chondroitin sulfate sodium (Chondroitin sulfate sodium), glucosamine (Glucosamine). 1 tablet contains chondroitin sulfate sodium - 500 mg, glucosamine hydrochloride - 500 mg. Excipients: disubstituted calcium sulfate - 230 mg, microcrystalline cellulose - 185 mg, croscarmellose sodium - 80 mg, stearic acid - 70 mg, magnesium stearate - 10 mg.

Shell composition: hydroxypropyl methylcellulose - 35 mg, titanium dioxide (E171) - 8.2 mg, triacetin - 6.8 mg.

Capsules based on cholesterol are represented by Structum, Teraflex advance, Teraflex, Chondroitin AKOS, Chondroitin, Kondronova.

All of the above drugs have the active ingredient: chondroitin sulfate sodium (Chondroitin sulfate sodium).

Gelatin capsules have excipients, such as talc, stearic acid, magnesium stearate, manganese sulfate.

Typically, the composition of the capsule shell includes titanium dioxide and gelatin.

Gels based on cholesterol are known, for example, the drug Chondroitin (Chondroitine, VERTEKS AO (Russia) - gel for external use 5%: tubes 30 g or 50 g, cans 30 g. Active ingredient:

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Chondroitin sulfate sodium (Chondroitin sulfate sodium). Gel for external use 5% - 1 g. Ingredients: chondroitin sulfate - 50 mg. Auxiliary substances are added to the gel: dimethyl sulfoxide;

propylene glycol; carbomer; trolamine; macrogol; edetat; methylparaben; nipagin; neroli and lavender oil; purified water.

Ointments based on cholesterol are represented by the following preparations: Mukosat ointment 5%, Artrafik ointment 5%, Chondroitin AKOS ointment 5%. Ointments contain sodium chondroitin sulfate and excipients: lactose, calcium stearate.

Injection forms are represented by lyophilizates and solutions.

Known solutions for intramuscular injection in ampoules: Drastop, Chondrogard, Injectran, Atravir-Incamfarm, Artogistan.

An example of a lyophilizate is Chondrolon, which is a lyophilized form of an injectable preparation based on the substance of the sodium salt of chondroitin sulfate.

All of the above drugs based on chondroitin sulfate (CS), whether injectable forms, gels, ointments, tablets or capsules for oral administration, having a chondroprotective effect, anti-inflammatory activity, acting mainly on inflammatory mediators, stimulate the synthesis of hyaluronic acid and proteoglycans and inhibit the action proteolytic enzymes.

In more detail, cholesterol preparations have a chondrostimulating, regenerating, anti-inflammatory and analgesic effect; have chondroprotective properties, enhance metabolic processes in hyaline and fibrous cartilage, subchondria bone; inhibit enzymes that cause degradation (destruction) of articular cartilage; stimulate the production of proteoglycans by chondrocytes; help reduce the release of inflammatory mediators and pain factors into the synovial fluid; suppress the secretion of leukotrienes and prostaglandins; slow down bone resorption and reduce calcium loss; accelerate the process of restoration of bone tissue; slow down the progression of osteoarthritis and osteochondrosis; contribute to the restoration of the articular bag and cartilaginous surfaces of the joints; prevent the collapse of connective tissue and normalize the production of joint fluid.

From the above data, it can be seen that cholesterol drugs in various forms are actively used for the treatment of arthrological diseases.

At the same time, for all the above drugs for the treatment of arthrological diseases based on cholesterol, a number of common technical problems are characteristic.

One such problem is microbial contamination. The causes of contamination of medicinal products are, in particular, inadequate conditions for obtaining, storing or transporting a medicinal product or components intended for its preparation. Raw materials of animal origin are among the most polluted.

Also, an increase in contamination may be associated with prolonged and / or improper storage of the drug. That is, there is a problem of reducing the shelf life of the drug, which, in turn, reduces the possibility of its effective use and requires additional efforts to dispose of an unused drug with an early expiration date.

Contamination leads not only to a decrease up to the complete absence of the necessary therapeutic effect, but also to the occurrence of inflammatory processes and the risk of infectious diseases during operation. Taking into account the fact that treatment with the drug involves long-term and repeated use of it, such risks can be very significant.

A separate problem of preparations obtained on the basis of natural organic raw materials is their potential allergenicity. The likelihood of allergic diseases with prolonged use of the drug also increases significantly.

As a rule, to combat pathogenic microorganisms in preparations of this kind, preservatives are used, which are various derivatives of benzene.

For example, the drug Mucosat, which is a solution for injection, includes the sodium salt of chondroitin sulfate - 8-12%, as a preservative - benzyl alcohol - 0.8-1.2% and pyrogen-free water - the rest, the pH of the drug solution is 6.5- 7.5.

This medicinal product contains benzyl alcohol as a preservative. However, this substance does not completely eliminate contamination, but only somewhat suppresses and, thus, slows down the growth of microorganisms, including microbes and fungi, and also dictates high requirements for the temperature regime of storage, failure to comply with which entails a significant decrease in the activity of the active substance. Thus, the problem with long-term storage persists.

In addition, benzyl alcohol exhibits in some cases (as side effects) the following toxic effects: respiratory failure, vasodilation, hypotension, convulsions and paralysis. It is also responsible for the development of gasping syndrome in children, characterized by respiratory failure, changes in motor functions, hypotension, renal failure, and thrombocytopenia. Obviously, with the systematic and prolonged use of the drug, which is typical for arthrological diseases, the risk of these negative consequences objectively increases.

Thus, the task of creating preparations of chondroitin sulfate that are stable during storage and retain their therapeutic efficacy during long-term storage and/or operation, while not exhibiting negative side effects associated with the presence of a relatively large amount of preservative, remains unresolved at present. .

The present invention is based on the technical problem of creating a product that is stable during storage and operation by effectively controlling the natural level of contamination and minimizing the risk of allergic reactions in the treatment of arthrological diseases.

Also, the present invention is based on the task of minimizing the content of the preservative, i.e. to achieve the stability of the product by including in its composition the minimum amount of a preservative, such as benzyl alcohol or sodium benzoate.

Disclosure of invention

The authors of the invention solve this problem by creating an agent for the treatment of arthrological diseases, which includes in its composition a pharmaceutically effective amount of the active substance - chondroitin sulfate and metallic or colloidal silver in a relative amount of from 0.1 to 10 mg/kg in terms of the mass of the active substance - chondroitin sulfate .

In a particular variant, in a relative amount from 0.2 to 8 mg/kg, and also from 0.3 to 6 mg/kg, calculated on the weight of the active substance.

In a private variant, in a relative amount of 0.4 to 5 mg/kg, and also from 0.5 to 4 mg/kg, and also from 0.2 to 3 mg/kg, and also from 0.15 to 2 mg/kg kg, as well as from 7 to 10 mg/kg, as well as from 4 to 8 mg/kg, calculated on the weight of the active substance.

In this case, the proposed agent, depending on the concentration of silver introduced into the composition, can either be free of a preservative (benzyl alcohol, sodium benzoate, or others with similar properties), or, if necessary, include it in a smaller amount.

The technical result, manifested when using the invention, is the stability of the agent for a long time and / or during operation due to the absence of microbial contamination, the preservation of its effectiveness and safety. It is important that this effect is manifested for various forms of the preparation containing this agent, which is of great importance not only for injection forms (solution, lyophilisate), but also for the storage of oral (tablet, capsule) and topical forms for the first opening of the package ( ointment, gel).

It is also important that when using the claimed agent, there are no negative effects associated with the presence of a relatively large amount of a preservative, including benzene derivatives. The risk of allergic reactions is reduced, which is due both to the exclusion or minimization of the preservative, and the stabilization of the active principle with silver in the stated concentrations.

It has been established that the addition of silver to the composition of the product in the indicated amount gives the maximum suppression of negative effects and avoids possible risks. These negative effects and risks include allergic reactions, as well as the toxic accumulation of silver in the organs and tissues of the body.

Thus, the proposed concentrations of silver unexpectedly turned out to be optimal for solving the problem.

Today, silver is actively used in the food, cosmetic and therapeutic industries. The popularity of silver compounds is due to their high antifungal, antibacterial and anti-inflammatory properties.

In medicine, such compounds are usually used as an independent active substance.

For example, in the patent of the Russian Federation 2268074 (published on January 20, 2006) a method for the treatment of rheumatoid arthritis is described by drug electrophoresis on the area of the affected joints according to the transverse technique. According to the invention, electrophoresis of a 1-2% solution of silver nitrate is carried out daily in the number of procedures of at least 7, while the first procedure is carried out with a duration of 15 minutes, and subsequent ones - 20 minutes. During electrophoresis with a 1-2% solution of silver nitrate, a depot (accumulation) of silver ions is formed in the upper layers of the skin above the affected joints, which provides a prolonged effect on the site of exposure. The concentration of the silver nitrate solution was selected empirically and, as observations have shown, is optimal for solving the problem. At a concentration of less than 1%, a slight effect is observed, and at a concentration of more than 2%, adverse reactions occur in the form of itching.

This method has shown effectiveness in rheumatoid arthritis and Bechterew's disease in terms of a significant reduction in pain, stiffness in the joints and spine, reduced swelling in the joints, positive dynamics of laboratory indicators of inflammation.

However, the use of silver for medical purposes has a number of limitations.

So, for example, the disadvantage of the method described above is the possibility of skin itching as a side effect due to the penetration of the drug through the skin.

When administered internally, silver nitrate (AgNO ₃ ) decomposes into pure silver, nitrogen dioxide and oxygen:

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2AgNO ₃ ^ 2Ag + 2NO2 + O ₂ , where pure silver itself is accumulated by the body and excreted only through the gastrointestinal tract.

The mass fraction of silver in silver nitrate AgNO ₃ is 63.5%, 2% solution of silver nitrate contains 1.27% pure silver.

In electrophoresis, 20-30 ml of a solution containing 254-381 mg of pure silver is usually used.

As mentioned above, the clinical effect of the drug is manifested when using a 1-2% solution of silver nitrate, in which the minimum dose of silver that enters the body is 127 mg.

It is known that for a human the toxic dose of silver is 50-60 mcg/l.

Consequently, the internal administration of the drug in the form of a solution of silver nitrate is fraught not only with allergic reactions, but also with a toxic accumulation of silver in excess of the daily dose, which can cause argyria.

It is known that leukocytes absorb silver and deliver it to the site of infection. And under the influence of silver, inflammation is removed: the metal and proteins form special compounds that can neutralize the action of most pathogenic microbes. Moreover, these compounds are capable of exerting a regenerating effect on tissues. Actually, this explains the beneficial effect of silver in any inflammation.

The active healing element of silver is its ions - Ag ⁺ . Silver ions easily interact with other ions, forming, as a rule, insoluble or poorly soluble compounds, which prevents the penetration of silver into a living organism. For example, silver ions readily bond with chloride anions, resulting in the formation of sparingly soluble silver chloride.

However, in an aqueous solution, silver ions retain their activity due to their stable stabilization in the form of hydrated silver. This is due to the property of a bipolar water molecule to form hydrogen bonds, as a result of which water molecules in the form of a shell can shield anions and cations from mutual influence.

In fact, fragments of the water molecule form a fairly stable shell around the silver ion, which keeps this ion in a free and active state. Therefore, even getting into the intestines, stabilized hydrated silver is not significantly affected by chloride ions and safely enters the body.

Also easily under the protection of water molecules, silver ions penetrate into skin tissues. Circulating in the bloodstream and liquid media of tissues, hydrated silver ions freely penetrate into pathogenic microbes, viruses and fungi through their outer shell, and then combine with the respiratory enzyme and, blocking the respiratory function, lead to their death. At the same time, pathogenic microorganisms cannot develop resistance to the destructive effect of silver ions, as they do with respect to antibiotics.

Known drug Colloidal silver in the form of an ionized solution in a 60 ml vial, which consists of the active ingredient: colloidal silver in the form of an ionized solution intended for internal and external use (Latin name: Silver colloidal, active ingredient: Colloidal silver (Silver colloidal), manufacturer : Nittany Pharmaceuticals/Vitaline, Nature's Sunshine Products (USA), Coral club (Germany) Used for rheumatoid arthritis.

In the treatment of the drug, the development of allergic reactions is also possible.

According to the World Health Organization, the ability to kill certain bacteria with certainty is observed at concentrations of silver ions above 150 μg / l, and at lower concentrations they only inhibit the reproduction of bacteria.

Therefore, the use of colloidal silver also has a clinical effect only when the toxic dose of silver is exceeded by 50-60 µg/l.

The least toxic metallic silver. Poorly soluble silver salts (for example, chloride) have low toxicity, but are also inactive in terms of bactericidal activity.

These data allow us to conclude that preparations containing silver can not only exhibit antibacterial activity, but also have technical problems: the difficulty of delivery without toxication of the body as a whole.

The authors of the present invention have created an agent containing an effective amount of chondroitin sulfate and silver, which makes it possible to provide the necessary therapeutic effect, avoid microbial contamination during manufacture, storage and operation, and at the same time minimize the negative effects associated with the presence of silver in the composition.

The amount of chondroitin sulfate sodium in the product can be from 50 to 2500 mg.

The amount of chondroitin sulfate sodium in the product can be from 2000 to 2500 mg.

The amount of chondroitin sulfate sodium in the product can be from 1500 to 2500 mg.

The amount of chondroitin sulfate sodium in the product can be from 1000 to 2500 mg.

The amount of chondroitin sulfate sodium in the product can be from 100 to 2000 mg.

The amount of chondroitin sulfate sodium in the product can be from 1500 to 2000 mg.

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The amount of chondroitin sulfate sodium in the product can be from 1000 to 2000 mg.

The amount of chondroitin sulfate sodium in the product can be from 500 to 2000 mg.

The amount of chondroitin sulfate sodium in the product can be from 300 to 1500 mg.

The amount of chondroitin sulfate sodium in the product can be from 1000 to 1500 mg.

The amount of chondroitin sulfate sodium in the product can be from 500 to 1500 mg.

The amount of chondroitin sulfate sodium in the product can be from 800 to 1500 mg.

The amount of chondroitin sulfate sodium in the product can be from 500 to 1200 mg.

The amount of chondroitin sulfate sodium in the product can be from 50 to 250 mg.

The amount of chondroitin sulfate sodium in the product can be from 50 to 100 mg.

The amount of chondroitin sulfate sodium in the product can be from 50 to 150 mg.

The amount of chondroitin sulfate sodium in the product can be from 50 to 300 mg.

The amount of chondroitin sulfate sodium in the product can be from 50 to 350 mg.

The amount of chondroitin sulfate sodium in the product can be from 100 to 400 mg.

The amount of chondroitin sulfate sodium in the product can be from 150 to 450 mg.

The amount of chondroitin sulfate sodium in the product can be from 250 to 1000 mg.

The agent of the present invention includes metallic or colloidal silver in a relative amount of 0.1 to 10 mg/kg, calculated on the weight of the active substance - chondroitin sulfate.

In a particular embodiment, silver is contained in a relative amount of 0.2 to 8 mg/kg, as well as from 0.3 to 6 mg/kg, based on the weight of the active substance.

In a particular embodiment, silver is contained in a relative amount of 0.4 to 5 mg/kg, and also from 0.5 to 4 mg/kg, and also from 0.2 to 3 mg/kg, and also from 0.15 to 2 mg/kg in terms of the mass of the active substance.

In a particular variant, silver is contained in a relative amount of 1 to 8 mg/kg, and also from 2 to 8 mg/kg, and also from 3 to 8 mg/kg, and also from 4 to 8 mg/kg, and also from 5 up to 8 mg/kg in terms of the mass of the active substance.

In a particular variant, silver is contained in a relative amount of 1 to 6 mg/kg, and also from 2 to 6 mg/kg, and also from 3 to 6 mg/kg, and also from 4 to 6 mg/kg, calculated on the mass of active substances.

In a particular embodiment, silver is contained in a relative amount of 1 to 5 mg/kg, as well as from 2 to 5 mg/kg, and also from 3 to 5 mg/kg, based on the weight of the active substance.

In a particular embodiment, silver is contained in a relative amount of 2 to 10 mg/kg, and also from 3 to 10 mg/kg, and also from 4 to 10 mg/kg, and also from 5 to 10 mg/kg, from 6 to 10 mg/kg, as well as from 8 to 10 mg/kg in terms of the mass of the active substance.

The mass ratio of sodium chondroitin sulfate to metallic or colloidal silver can be 1:1x10' ⁷ - 1:1x10'5.

As part of the product, metallic or colloidal silver can be used.

More specifically, silver gels or sols may be used in the agent.

The size of the silver particles used can be 1-100 µm - 1-100 nm.

Colloidal silver can be used, for example, AgBion-2 colloidal nanosilver concentrate [http://rps61.ru/good/65] or KND-S colloidal silver concentrates [http://ultrasilver.ru]

The agent of the present invention may be in liquid or solid form.

The tool can be used for oral, injection or topical application.

Forms intended for topical use include creams, gels and ointments.

Oral forms include tablets and capsules.

Injectable forms include lyophilisates and injectable solutions.

Auxiliary components are selected depending on the form of manufacture.

The tool, made in the form of a cream, gel or ointment, may contain one or more excipients selected from the group: dimethyl sulfoxide; propylene glycol; carbomer; trolamine; macrogol; edetat; neroli and lavender oils; lanolin; petrolatum.

The tool, made in the form of a tablet, may additionally contain glucosamine hydrochloride.

In a private embodiment, the ratio of chondroitin sulfate sodium to glucosamine hydrochloride in a tablet is 1:1.

The tablet may additionally contain the following excipients, taken in relation to sodium chondroitin sulfate:

disubstituted calcium sulfate -1:0.46, microcrystalline cellulose - 1:0.37, croscarmellose sodium - 1:0.16, stearic acid - 1:0.14, magnesium stearate - 1:0.02.

In a private embodiment, the tablet may be film-coated, in which case the tablet shell has the following composition, taken in relation to chondroitin sulfate sodium:

- 5 041409 hydroxypropylmethylcellulose - 1:0.07, titanium dioxide (E171) - 1:0.016, triacetin - 1:0.014.

The agent, made in the form of a capsule, may additionally contain excipients selected from the group: talc, stearic acid, magnesium stearate, manganese sulfate.

The composition of the capsule shell may include titanium dioxide and gelatin.

The tool, made in the form of a solution, contains water in its composition.

More specifically, the product contains distilled or pyrogen-free water.

The finished product may include in its composition a certain amount of preservatives, such as benzyl alcohol and sodium benzoate or the like.

The specific ratios of the components are determined by the form of manufacture and the severity of the disease.

The claimed agent is intended for the treatment of arthrological diseases.

Such diseases include degenerative-dystrophic diseases of the joints and spine: osteoarthritis of peripheral joints; intervertebral osteochondrosis and osteoarthritis, rheumatoid arthritis, gout, reactive arthritis, spondyloarthropathies, arthritis in systemic lupus erythematosus, dermatomyositis and other systemic diseases, psoriatic arthritis, post-traumatic damage to articular cartilage, ligaments, menisci.

Image list

Fig. 1: radiograph of the knee joints of rabbits of groups I and V;

fig. 2: radiograph of the knee joints of rabbits treated intraarticularly in the right (R) joint with papain and preparations A and B. The left (L) joint is intact;

fig. 3: X-ray of the knee joints of rabbits of group IV, treated intra-articularly with papain and the control drug (comparison) in the right (R) joint;

fig. 4: articular cartilage of the rabbit knee joint of group I;

fig. 5: synovium of the right knee joint of a rabbit of group I;

fig. 6: articular cartilage of the rabbit knee joint after intramuscular administration of Preparation A, Preparation B, saline C.

Implementation of the invention

The following are examples of the preparation of various drugs based on the claimed means of chondroitin sulfate.

Example 1

A gel preparation containing sodium chondroitin sulfate, purified water and additionally containing metallic or colloidal silver from 0.10 to 10 mg/kg can be obtained using various excipients.

1. The preparation in the form of a gel is obtained by first forming solution A: by mixing purified water and carbomer, which is taken in a volume of 3% of the total mass. Then solution B is formed: by dissolving methyl parahydroxybenzoate in ethanol, where ethanol and methyl parahydroxybenzoate are taken in a volume of 10 and 0.1% of the total mass, respectively. To obtain solution B, the following are mixed when heated to a temperature of 65-75 ° C in purified water (wt. / vol.%):

g chondroitin sulfate, dimethyl sulfoxide - 7.5;

propylene glycol - 7.5;

trolamine (triethanolamine) - 3;

macrogol glycerylhydroxystearate - 0.4;

disodium edetate - 0.1.

After that, solutions A, B, C are constantly stirred for several hours until a helium mass is formed, adding metallic or colloidal silver 0.0005% of the total mass of the total mass. If necessary, add flavors (wt./vol.%):

lavender oil - 0.1;

orange blossom oil (neroli oil) - 0.05.

The resulting finished composition of the drug in the form of a gel contains (wt./vol.%):

chondroitin sulfate - 5;

ethanol - 10;

dimethyl sulfoxide - 7.5;

propylene glycol - 7.5;

carbomer - 3;

trolamine (triethanolamine) - 3;

macrogol glycerylhydroxystearate - 0.4;

disodium edetate - 0.1;

methyl parahydroxybenzoate (methylparaben, nipagin) - 0.1;

lavender oil - 0.1;

orange blossom oil (neroli oil) - 0.05;

- 6 041409 metallic or colloidal silver - 0.0005;

purified water - the rest up to 100.

2. Solution A is obtained by adding 10 g of carbomer to 120 ml of purified water, followed by stirring. To obtain solution B, 0.2 g of methyl parahydroxybenzoate is dissolved in 60 g of ethanol. Solution B is prepared by stirring with heating to a temperature of 70°C in 181.6 ml of purified water, 2 g of chondroitin sulfate, 2 g of glucosamine sulfate, 4 g of methylsulfonylmethane, 0.04 g of sodium metabisulphite, 20 g of propylene glycol. After that, solutions A, B, C are constantly stirred for several hours until a helium mass is formed, adding metallic or colloidal silver 0.0002% of the total mass. If necessary, add a perfume in the amount of 0.2 g. The resulting finished composition of the drug in the form of a gel contains (wt./vol.%):

chondroitin sulfate - 5;

glucosamine sulfate - 0.5;

methylsulfonylmethane - 1.0;

sodium metabisulphite - 0.01;

methyl parahydroxybenzoate - 0.05;

propylene glycol - 5.0;

ethanol - 15.0;

carbomer - 2.5;

fragrance - 0.05;

metallic or colloidal silver - 0.0002;

purified water - the rest up to 100.

Example 2

The preparation in the form of an ointment, including sodium chondroitin sulfate, purified water and additionally containing metallic or colloidal silver from 0.1 to 10 mg/kg, can also be obtained using various excipients.

1. Medical vaseline in an amount of 37 wt.%, anhydrous lanolin in an amount of 15 wt.%, stearic acid in an amount of 3 wt.%, pentol in an amount of 5 wt.% are melted at a temperature of (70 ± 2) ° C, mixed, then cooled to a temperature of (55±2)°C (ointment base). Chondroitin sulfate in an amount of 5 wt.% is dissolved in 20 wt.% distilled water. Diclofenac sodium in an amount of 5 wt.% is dissolved in 10 wt.% dimethyl sulfoxide. In solutions of sodium diclofenac in dimethyl sulfoxide and chondroitin sulfate in water, metallic or colloidal silver 0.00005% of the total mass is added with stirring, the resulting substance is added to the ointment base, homogenized, and then transferred to packaging. Get the finished ointment having the composition (wt./vol.%):

chondroitin sulfate sodium (in terms of dry matter) - 5;

dimethyl sulfoxide - 10;

lanolin - 0.15;

vaseline - 0.5;

metallic or colloidal silver - 0.00005;

purified water - the rest up to 100.

2. Medical vaseline in an amount of 37 wt.%, anhydrous lanolin in an amount of 15 wt.%, stearic acid in an amount of 3 wt.%, pentol in an amount of 5 wt.% are melted at a temperature of (70 ± 2) ° C, mixed, then cooled to a temperature of (55±2)°C (ointment base). Chondroitin sulfate in an amount of 5 wt.% is dissolved in 20 wt.% distilled water. Diclofenac sodium in an amount of 5 wt.% is dissolved in 10 wt.% dimethyl sulfoxide. In solutions of sodium diclofenac in dimethyl sulfoxide and chondroitin sulfate in water, metallic or colloidal silver is added with stirring to 0.0002% of the total mass, the resulting substance is added to the ointment base, homogenized, and then transferred to packaging. Get the finished ointment having the composition (wt./vol.%):

chondroitin sulfate sodium (in terms of dry matter) - 5;

dimethyl sulfoxide - 10;

lanolin - 0.15;

vaseline - 0.5;

metallic or colloidal silver - 0.0002;

purified water - the rest up to 100.

Example 3

The drug contains 95-120 mg of lyophilized powder of Na-salt of chondroitin sulfate, which is obtained from the sodium salt of chondroitin sulfate dissolved in pyrogen-free injection water.

The injection solution prepared from the said lyophilizate using pyrogen-free water contains Na-salt of chondroitin sulfate in a therapeutic dosage of 9.0-11.5 wt.%.

The preparation can be obtained, for example, in the following way.

A substance with a sodium salt content of chondroitin sulfate of 96.2-97.1% is dissolved in 1000 ml of pyrogen-free injection water, the pH is set from 6.0 to 7.5 with sodium hydroxide solution,

- 7 041409 after which, in relation to the resulting mixture, the process of ionization with silver ions Ag ⁺ is carried out to the level

0.01-1.0 mg/l of the mixture, then sterilizing filtration is carried out, poured into 1 ml ampoules and sublimation is carried out in a known manner, the ampoules with the lyophilized drug are sealed.

1. 104 g of a substance containing 96.2% sodium salt of chondroitin sulfate is dissolved in 1000 ml of pyrogen-free injection water, pH is adjusted from 6.0 to 7.5 with sodium hydroxide solution. Then, in relation to the resulting mixture, the process of ionization with silver ions Ag ⁺ to a level of 0.10 mg/l of the mixture is carried out. Then the process of sterilizing filtration is carried out, poured into 1 ml ampoules and sublimation is carried out in a known manner. Ampoules with lyophilized drug are sealed. 103 mg of lyophilized powder is obtained in one ampoule, which corresponds to 99.7 mg of the sodium salt of chondroitin sulfate, the residual moisture is the rest, which, when dissolved in 1 ml of injection water, will give a solution with an active substance content of 9.97%.

2. 103 g of a substance containing 96.2% sodium salt of chondroitin sulfate is dissolved in 1000 ml of pyrogen-free injection water, the pH is adjusted from 6.0 to 7.5 with sodium hydroxide solution. Then, in relation to the resulting mixture, the process of ionization with silver ions Ag ⁺ to a level of 0.50 mg/l of the mixture is carried out. Then the process of sterilizing filtration is carried out, poured into 1 ml ampoules and sublimation is carried out in a known manner. Ampoules with lyophilized drug are sealed. 103 mg of lyophilized powder is obtained in 1 ampoule, which corresponds to 99.7 mg of the sodium salt of chondroitin sulfate, the residual moisture is the rest, which, when dissolved in 1 ml of injection water, will give a solution with an active substance content of 9.97%.

Example 4

Tablet containing chondroitin sulfate.

The composition in the form of a tablet can be prepared as follows: weighed chondroitin sulfate sodium, glucosamine hydrochloride in a ratio of 1:1, colloidal silver from 0.10 to 10 mg/kg are mixed in a drum-type mixer. The resulting mixture is molded and pressed, then dried to the required residual moisture and calibrated. Then they are enclosed in a hard shell, which can have a composition taken in relation to sodium chondroitin sulfate: hydroxypropyl methylcellulose - 1: 0.07, titanium dioxide (E171) - 1: 0.016, triacetin - 1: 0.014 mg. Also, the tablet may additionally contain excipients taken in relation to sodium chondroitin sulfate: disubstituted calcium sulfate - 1:0.46, microcrystalline cellulose - 1:0.37 mg, croscarmellose sodium - 1:0.16, stearic acid - 1: 0.14, magnesium stearate - 1:0.02.

1. Tablet containing chondroitin sulfate.

The composition in the form of a tablet can be prepared as follows: weighed chondroitin sulfate sodium - 250 mg, colloidal silver - 0.6 mg/kg; calcium stearate - 1:0.019; crospovidone - 1:0.05; povidone-K30 - 1:0.04; microcrystalline cellulose - 1:0.38; magnesium bicarbonate pentahydrate - 1:0.429 is mixed in a drum mixer. The resulting mixture is molded and pressed, then dried to the required residual moisture and calibrated.

2. Tablet containing chondroitin sulfate and glucosamine hydrochloride.

The composition in the form of a coated tablet can be prepared as follows: chondroitin sulfate sodium, glucosamine hydrochloride in a ratio of 1:1.25; colloidal silver - 0.2 mg/kg; methylsulfonylmethane - 1:0.75. Can also be added: cellulose - 1:0.50; calcium hydrophosphate dihydrate - 1:0.3; stearic acid - 1:0.18; croscarmellose sodium - 1:0.125; magnesium stearate - 1:0.025; colloidal silicon dioxide - 1:0.01. The resulting mixture is molded and pressed, then dried to the required residual moisture and calibrated. Then they are enclosed in a shell, which may have the following composition, taken in relation to the amount of chondroitin sulfate: hypromellose - 1:0.04; polydextrose. - 1:0.02; titanium dioxide - 1:0.0001; talc -1:0.02; maltodextrin - 1:0.0117; triglycerides - 1:0.0015; E110 - 1:0.0023.

The above examples show that the preparation of preparations can be implemented using various excipients that do not affect the final result, since the functional role of the agent for the treatment of arthrological diseases and the achievement of the claimed technical result is determined by the active substance based on sodium chondroitin sulfate, to which a metallic or colloidal silver from 0.1 to 10 mg/kg.

Microbiological control of contamination in solid form was carried out. In particular, after 6 months and 12 months of storage of the finished form, no significant changes were detected.

Example 5

Evaluation of resistance to microbial contamination.

1. Cream, gel, ointment.

The effect was detected on the bacteria Staphylococcus aureus and E. Coli.

When growing Staphylococcus aureus, a nutrient medium based on yolk-salt agar and growth stimulants was used.

The growth of E. coli biomass was carried out in a nutrient medium at 37°C and 150 rpm for 24 h.

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In ointments and gels based on cholesterol and silver obtained according to the invention (examples 1, 2), a separate inoculation of the obtained cultures of Staphylococcus aureus and E. Coli was carried out.

As reference drugs, ointments and gels of a similar composition containing silver at concentrations of 0.040 and 0.070 mg/kg were used.

The growth of bacterial colonies was not observed for several hours in ointments and gels with a content of colloidal silver in it from 0.20 mg/kg and above.

With a decrease in the level of colloidal silver (comparative drugs), the growth of bacterial colonies occurred immediately and had a progressive effect.

2. Solution.

The effect was detected on the bacteria Staphylococcus aureus and E. Coli.

When growing Staphylococcus aureus, a nutrient medium based on yolk-salt agar and growth stimulants was used.

The growth of E. coli biomass was carried out in a nutrient medium at 37°C and 150 rpm for 24 h.

In solutions based on cholesterol and silver obtained according to the invention (example 3), a separate inoculation of the obtained cultures of Staphylococcus aureus and E. Coli was carried out.

As reference drugs, CS solutions of the same composition containing silver at concentrations of 0.040 and 0.070 mg/kg were used.

The growth of bacterial colonies was not observed for several hours in solutions saturated with silver ions Ag ⁺ from 0.20 mg/kg and above. With a decrease in the level of silver ions, the growth of bacterial colonies occurred immediately and had a progressive effect.

Example 6

Evaluation of resistance to microbial contamination.

1. Gel.

Studies have been conducted on the following drugs.

drug A.

Gel containing, mg: chondroitin sulfate - 100; ethanol - 1000; dimethyl sulfoxide - 75; propylene glycol - 75; carbomer - 300; trolamine - 300; macrogol glycerylhydroxystearate - 40; disodium edetate - 10; methyl parahydroxybenzoate - 10; lavender oil - 10; orange blossom oil - 5; colloidal silver - 0.0001; water - up to 10000.

Comparator A1.

Gel containing, mg: chondroitin sulfate - 100; ethanol - 1000; dimethyl sulfoxide - 75; propylene glycol - 75; carbomer -300; trolamine - 300; macrogol glycerylhydroxystearate - 40; disodium edetate - 10; methyl parahydroxybenzoate - 10; lavender oil - 10; orange blossom oil - 5; water - up to 10000.

drug B.

Gel containing, mg: chondroitin sulfate - 300; ethanol - 1000; dimethyl sulfoxide - 75; propylene glycol - 75; carbomer - 300; trolamine - 300; macrogol glycerylhydroxystearate - 40; disodium edetate - 10; methyl parahydroxybenzoate - 10; lavender oil - 10; orange blossom oil - 5; colloidal silver - 0.0015; water - up to 10000.

Comparator B1.

Gel containing, mg: chondroitin sulfate - 300; ethanol - 1000; dimethyl sulfoxide - 75; propylene glycol - 75; carbomer -300; trolamine - 300; macrogol glycerylhydroxystearate - 40; disodium edetate - 10; methyl parahydroxybenzoate - 10; lavender oil - 10; orange blossom oil - 5; water - up to 10000.

drug B.

Gel containing, mg: chondroitin sulfate - 500; ethanol - 1000;

dimethyl sulfoxide - 75; propylene glycol - 75; carbomer - 300; trolamine - 300; macrogol glycerylhydroxystearate - 40; disodium edetate - 10; methyl parahydroxybenzoate - 10; lavender oil - 10; orange blossom oil - 5; colloidal silver - 0.0035; water - up to 10000.

Comparator B1.

Gel containing, mg: chondroitin sulfate - 500; ethanol - 1000; dimethyl sulfoxide - 75; propylene glycol - 75; carbomer -300; trolamine - 300; macrogol glycerylhydroxystearate - 40; disodium edetate - 10; methyl parahydroxybenzoate - 10; lavender oil - 10; orange blossom oil - 5; water - up to 10000.

As comparison drugs, ointments and gels of a similar composition, not containing colloidal silver, were used.

The effect was detected on the bacteria Staphylococcus aureus and E. Coli.

When growing Staphylococcus aureus, a nutrient medium based on yolk-salt agar and growth stimulants was used.

The growth of E. coli biomass was carried out in a nutrient medium at 37°C and 150 rpm for 24 h.

- 9 041409

In all preparations, a separate sowing of the obtained cultures was carried out.

The absence or presence of bacterial colonies was recorded throughout the experiment every 30 min. The duration of the experiment is 8 hours.

A relationship was established between the concentration of colloidal silver and the growth of bacterial colonies.

In comparison preparations, bacterial growth was noted 30 min after the start of the experiment.

At a concentration of colloidal silver of 1 mg per 1 kg of chondroitin sulfate, bacterial growth was observed 3 hours after the start of the experiment.

At a concentration of colloidal silver of 5 mg per 1 kg of chondroitin sulfate, bacterial growth was observed 6 hours after the start of the experiment.

At a concentration of colloidal silver of 7 mg per 1 kg of chondroitin sulfate, bacterial growth was noted 5 hours after the start of the experiment.

Thus, according to the results of the experiment, a relationship was established between the level of contamination and the concentration of silver in the preparation.

Example 7

Studies have been conducted on the following drugs.

drug A.

Solution of the following composition:

Chondroitin sulfate - 100 mg; benzyl alcohol - 9 mg, silver proteinate - 0.0001 mg; water for injection - up to 1 ml.

drug A1.

Solution of the following composition:

Chondroitin sulfate - 100 mg; benzyl alcohol - 9 mg; water for injection - up to 1 ml.

drug B.

Solution of the following composition:

Chondroitin sulfate - 150 mg; benzyl alcohol - 9 mg, silver proteinate - 0.00015 mg; water for injection - up to 1 ml.

Preparation B1.

Solution of the following composition:

Chondroitin sulfate - 150 mg; benzyl alcohol - 9 mg; water for injection - up to 1 ml.

drug B.

Solution of the following composition:

Chondroitin sulfate - 100 mg; benzyl alcohol - 9 mg, silver proteinate - 0.0002 mg; water for injection - up to 1 ml.

drug B1.

Solution of the following composition:

Chondroitin sulfate - 200 mg; benzyl alcohol - 9 mg; water for injection - up to 1 ml.

As reference drugs, solutions of cholesterol of a similar composition, not containing silver proteinate, were used.

The effect was detected on the bacteria Staphylococcus aureus and E. Coli.

When growing Staphylococcus aureus, a nutrient medium based on yolk-salt agar and growth stimulants was used.

The growth of E. coli biomass was carried out in a nutrient medium at 37°C and 150 rpm for 24 h.

In all preparations, a separate sowing of the obtained cultures was carried out.

The degree of contamination (growth of bacterial colonies) was measured for 12 hours every 30 minutes.

The following results are obtained:

Progressive growth of bacterial cultures was observed in reference preparations A1, B1, C1 30 min after the start of the experiment.

In preparation A - 5 hours after the start of the experiment.

In preparation B - 7.5 hours after the start of the experiment.

In preparation B - 10 hours after the start of the experiment.

As a result of the experiment, a relationship was established between the concentration of silver proteinate in the studied preparations and the growth of bacterial cultures.

Example 8

The effectiveness of chondroitin sulfate preparations.

1. The effectiveness of products in the form of a gel or ointment, including sodium cholesterol, purified water, metallic or colloidal silver from 0.1 to 10 mg/kg, was tested on rabbits.

The studied samples were applied to animals several times. The activity was assessed on the model of destruction, which was caused by 6 injections of papain with an interval of 5 days into the cavity of the right knee joint at a dose of 50 mg per injection (1 ml of a 5% solution) - I, II, III, IV, V groups of 10 rabbits in each.

- 10 041409

Experimental animals on the knee joint with light rubbing daily for 15 days were applied:

for groups I and II - the studied samples of ointment (concentration of colloidal silver 0.04 mg/l) and gel (concentration of colloidal silver 0.1 mg/l), respectively;

for group III - chondroitin sulfate ointment, not containing metallic or colloidal silver, containing benzyl alcohol in its composition;

for the control group IV - isotonic sodium chloride solution.

In another control group V - nothing was applied.

The severity of edema was assessed by measuring its thickness before and after administration of papain.

When applied on the skin in groups I, II, III after 15 days, there was a decrease in the severity of edema of the knee joint in comparison with the control groups by 2.1 times.

The severity of the anti-inflammatory effect of the studied agents (ointment and gel) according to the invention and the silver-free ointment was observed and did not appear in the control groups IV, V.

With a longer observation, it was noted that groups I and II showed more active regeneration than group III.

In particular, out of 10 rabbits of each of groups I, II, III, 9 out of 10 in groups I, II after 35 days recorded complete regeneration of bone tissue, and in group III only 7 out of 10.

Thus, the claimed agents according to the invention showed anti-inflammatory activity, normalization of metabolic processes, as well as more effective regeneration of bone tissue than silver-free cholesterol ointment.

The presence of allergic reactions in animals was determined visually. In 7 rabbits from group II and 6 rabbits from group II, redness at the injection site and skin itching were observed. In group III, redness was observed in one animal, itching was not observed.

2. The effectiveness of injection forms.

The effectiveness of similar forms of silver-containing cholesterol intended for injection was studied and compared with the effect of injections of a cholesterol-containing preparation containing benzyl alcohol and not containing silver.

Namely, the following tools were used:

A) an agent for the treatment of arthrological diseases, which is a drug in the form of a lyophilized powder of Na-salt of chondroitin sulfate and residual moisture containing silver ions Ag ⁺ 0.1 mg/ml;

B) an agent for the treatment of arthrological diseases, which is a preparation in the form of a solution for intramuscular injection, including Na-salt of chondroitin sulfate from 4 to 12 wt.%, purified water, containing silver ions Ag ⁺ 0.2 mg / ml;

B) control preparation: an agent for the treatment of arthrological diseases, which is a preparation in the form of a solution for intramuscular injection, including Na-salt of chondroitin sulfate from 4 to 12 wt.%, benzyl alcohol, purified water.

At the same time, the tested injection solution was prepared once, stored under standard conditions, and administered, including after storage. So the drug was administered after 12 and 25 weeks of storage. At the same time, according to the results of a biochemical study of connective tissue components by spectrophotometry, no difference was found in the use of the drug immediately after its manufacture and after its storage (under standard conditions for 12 and 25 weeks). Standard control methods were used.

The study of preparations A and B was tested on 70 white rats of different sexes and an initial weight of 250-300 g. The animals were divided into 5 groups of 14 animals each.

The drug according to variants of the invention A and B was administered intramuscularly at doses of 0.25 and 0.5 g/kg and intravenously at doses of 0.1 and 0.25 g/kg.

Experimental batches of the dosage form of the drug were used. According to option A, the solution for administration was prepared on the basis of a lyophilized powder obtained according to the claimed invention.

Control animals received the reference drug at the same doses intravenously and intramuscularly.

The effect of the drug on the course of the inflammatory process was assessed by the change in the magnitude of the inflammatory edema of the limb (oncometrically), by the content of leukocytes and eosinophils in the blood.

The strength of the skin scar in rats was evaluated by two indicators - the force that causes the scar to break, and the relative elongation at break, which characterizes the elasticity of the tissue.

The effect of drugs on wound healing was studied on a model of a flat-bone skin wound.

Pathological and histochemical studies of the joints were performed randomly.

The material (synovium, articular cartilage of the knee joint) was fixed, embedded in paraffin, stained with hematoxylin-eosin, and total GAGs were determined with safranin-T. For separate detection of cholesterol, alcian blue staining was used on acetate buffer (pH-5.8) with different contents of MgCl ₂ (0.4 M MgCl ₂ when reacting to cholesterol).

The GAG content was assessed visually in points. Thus, the absence of staining with safranin was considered as subthreshold (minimum) for this dye (-). The content of GAGs is focal, light red staining - as a low content of GAGs (+), diffuse light red - as moderate (++), diffuse bright red - as pronounced (+++). No alcian blue staining - (-), pale blue staining - as low (+), blue - as moderate (++), bright blue - as significant (+++).

Destruction was caused by 6 injections of papain.

Rats of I, II, III, IV groups were intravenously injected with papain at 10 mg/kg (1% solution) - 4 injections with an interval of 3-4 days.

Animals of the V (control) group received physiological saline at the same time.

A day after the last injection of papain, the rats of group II began to receive intramuscular injection of preparation A, 0.5 ml daily for a month. Rats of group III received intramuscularly preparation B at the same time, 0.5 ml. Rats of group IV received intramuscularly at the same time a control preparation of 0.5 ml.

At the end of the experiment, the rats were killed. The costal cartilages were separated, cleaned of soft tissues, dried with acetone, and the total GAG content was determined in them according to the method of D.V. Kosyagin. [Kosyagin D.V. - Issues of medical chemistry, 1982, 28, 4, p. 68-71].

The results of the studies showed that with the introduction of papain, the content of GAG in the costal cartilages of rats somewhat decreases (normally it is 14.25 ± 2.7 mg of uronic acid per 1 g of dry tissue), in untreated rats of group I, the decrease in GAG was 15% (12 .11±2.4 mg/g).

With the introduction of drug B, the content of GAG increased and was above the norm to the level of 18.03±2.23 mg/g.

In rats treated with preparation A, the GAG content also turned out to be higher than in control animals (18.72±2.11 mg/g).

In rats treated with the control preparation, the content of GAGs also turned out to be higher than in control animals (17.14±2.08 mg/g).

Thus, the results of experiments on rats showed that drug A and drug B have a chondroprotective effect in case of damage to cartilage tissue by papain, simulating the accumulation of GAG in the costal cartilages of rats slightly better than the control drug.

Evaluation of the effect of drugs on destructive changes in the joint in rabbits.

In the experiments, 35 male chinchilla rabbits weighing 2.1-2.6 kg were used. Destruction was caused by 6 injections of papain with an interval of 5 days into the cavity of the right knee joint at a dose of 50 mg per injection (1 ml of a 5% solution) - groups I, II, III, IV, 7 rabbits each. The group of rabbits, in relation to which the natural increase in the circumference of the edema of the joint was recorded, was injected intra-articularly with 1.0 ml of physiological solution - group V (7 rabbits).

Within 2 months, rabbits of group I were injected with saline, group II - preparation A, 1.0 ml 3 times a week, group III - preparation B, 1.0 ml 3 times a week, group IV (control) - 1, 0 ml of the reference drug.

The drugs were administered intramuscularly. Thus, the chondroprotective effect of preparations A and B and the control preparation were studied in order to evaluate the therapeutic effect of the influence of the silver + cholesterol complex and compare it with the preparation based on cholesterol (Mukosat), which also proved effective.

The circumference of the joint edema increased after papain injections. By the end of the second month of the experiment, the joint circumference in rabbits of group I (without treatment) was significantly increased (more than 18 mm). In groups II, III, who received drugs A and B, the edema was slightly less - 6.2±1.1 mm and 6.3±1.2 mm, respectively, compared with group IV, which received the reference drug (8.1± 1.3 mm).

In rabbits of group I, who received only papain, by the end of 2 months, the GAG content increased to 12.6 mg/ml, which indicated the destruction of cartilage tissue. In animals treated with drugs A, B and control, the level of GAG was significantly lower (6.56, 6.88 and 7.73 mg/ml, respectively), from which it was concluded that destructive processes were less pronounced. The condition of group V was expressed within the increment of 3.02 mg/ml.

Pathological changes in the joints of rabbits of group I (without treatment) were recorded during X-ray examination.

In FIG. 1 shows a radiograph of the knee joints of rabbits of groups I and V (two months of experience). Injections into the right (R) joint. The left (L) joint is intact.

A - Rabbit No. 1 - Group V. XS solution. Both joints are unchanged.

B - Rabbit No. 2 - group I without treatment (intra-articular papain); P - signs of arthritis-arthrosis, destructive changes in the condyles of the femur; L - no change.

B - Rabbit No. 3 - group I without treatment (papain intra-articular); P - synovitis; L - no change.

In FIG. Figure 2 shows a radiograph of the knee joints of rabbits treated intraarticularly with papain and preparations A and B in the right (R) joint. The left (L) joint is intact. Experience 2 months.

A - Rabbit No. 4 - II group. Both joints are unchanged.

- 12 041409

B - Rabbit No. 5 - IV group. Both joints are unchanged.

In FIG. 3 shows a radiograph of the knee joints of rabbits of group IV, which received intra-articular papain and the control drug (comparison) in the right (R) joint. The left joint (L) is intact.

Experience 2 months.

A - Rabbit No. 6 - P - arthritis, osteoporosis; L - no change.

B - Rabbit No. 7 - both joints are unchanged.

General data on the results of observations by the end of 2 months are shown in the table.

Type of destruction The number of rabbits in which destruction was revealed by the end of the experiments I group II group III group IV group Arthritis 2 0 1 2 Arthrosis 3 0 0 0 Synovitis 2 0 0 0 Osteoporosis 3 0 0 2

The table shows that in group I, destructive changes persisted by the 2nd month in each rabbit. Most of the rabbits in this group had pronounced complex signs of arthritis and arthritis-osteoporosis.

In groups II and III, treated with preparations A and B, only one rabbit from group III showed signs of arthritis. In group IV rabbits treated intraarticularly with papain and intramuscularly, a good therapeutic effect was also observed, and only two rabbits had arthritis and osteoporosis at the same time. In all other rabbits of groups II, III, IV, changes in the joints were not detected.

According to the results of the study, it can be concluded that the introduction of drugs A and B prevented the development of pathological changes in the joints, as well as the use of the control drug of comparison, but the preventive effect of drugs A and B is slightly better than that of the control based on cholesterol.

The positive effect of the action of preparations A and B was also confirmed by pathomorphological studies.

An autopsy of rabbits of group I showed that each of them had pronounced signs of destruction of the articular cartilage in the right joint (see Figs. 4 and 5).

In FIG. 4 shows the articular cartilage of the knee joint of a group I rabbit.

A - After intra-articular injection of papain solution. Usurs are visible, penetrating to the middle of the intermediate layer. Proliferation of chondrocytes alternates with areas of devastation. There is a decrease in GAG. Staining with safranin - T.10x6.3.

B - Control. The histological structure of the cartilage is preserved. The amount of GAG is normal. Staining with safranin - T.10x6.3.

In FIG. 5 shows the synovial membrane of the right knee joint of a group I rabbit.

A - After intra-articular administration of papain. Severe hyperplasia of the synovial membrane, synoviocytes are large, located loosely, form 5-5 layers. In the subsynovial layer, the proliferation of histicites, fibroblasts and diffuse macrofocal plasmacytic infiltration are visible. Hem.-eosin 25x6.3.

B - Control - normal structure.

At autopsy of rabbits II, III (see Fig. 6), the surface of the articular cartilage was smooth and shiny.

In FIG. 7 shows the articular cartilage of the rabbit knee joint after intramuscular injection of preparation A, preparation B, saline (C).

A and B show increased proliferation of chondrocytes with a significant increase in GAG in the matrix.

B - The histological structure is preserved. Group V. Safranin - T.25x6.6.

In two rabbits of group IV, who received control Mukosat simultaneously with papain intramuscularly, a small amount of serous exudate was detected in the joint cavity. And all the rabbits of this group had a smooth and shiny surface of the articular cartilage.

These studies show that preparations A and B have the best effect on bone tissue regeneration, while biochemically and immunologically, manifestations of allergic reactions have not been recorded in these groups of animals.

Example 9

The effectiveness of chondroitin sulfate preparations.

Studies have been conducted on the following drugs.

drug A.

An agent for the treatment of arthrological diseases, which is a preparation in the form of a lyophilized powder containing 100 mg of Na-salt of chondroitin sulfate, silver ions Ag ⁺

0.0001 mg and residual moisture 7-10 mg.

drug A1.

An agent for the treatment of arthrological diseases, which is a drug in the form of a lyophilized powder containing 100 mg of Na-salt of chondroitin sulfate and residual moisture of 7-10 mg.

drug B.

An agent for the treatment of arthrological diseases, which is a preparation in the form of a lyophilized powder containing 100 mg of Na-salt of chondroitin sulfate, silver ions Ag ⁺ 0.0005 mg and residual moisture 7-10 mg.

Preparation B1.

An agent for the treatment of arthrological diseases, which is a drug in the form of a lyophilized powder containing 100 mg of Na-salt of chondroitin sulfate and residual moisture of 7-10 mg.

drug B.

An agent for the treatment of arthrological diseases, which is a preparation in the form of a lyophilized powder containing 100 mg of Na-salt of chondroitin sulfate, silver ions Ag ⁺ 0.0007 mg and residual moisture 7-10 mg.

drug B1.

An agent for the treatment of arthrological diseases, which is a drug in the form of a lyophilized powder containing 100 mg of Na-salt of chondroitin sulfate and residual moisture of 7-10 mg.

The silver ionization process has been used to increase the stability of the solution, since in aqueous solution silver ions retain their activity better due to their stable stabilization as hydrated silver.

The tested injection solution was prepared once, stored under standard conditions, and administered, including after storage.

The drug was administered after 12 and 25 weeks of storage. According to the results of a biochemical study of connective tissue components by spectrophotometry, no difference was found in the use of the drug immediately after its manufacture and after its storage (under standard conditions for 12 and 25 weeks).

Standard control methods were used.

The study of drugs was tested on 60 white rats of different sexes and an initial weight of 250300 g.

The rats were divided into three groups of 20 animals each.

Group No. 1 - trials of preparation A and reference preparation A1.

Group No. 2 - trials of preparation B and comparison preparation B1.

Group 3 - trials of preparation B and reference preparation B1.

Each group was divided into four subgroups (I, II, III, IV).

Experimental batches of the dosage form of the drug were used.

The effect of the drug on the course of the inflammatory process was assessed by the change in the magnitude of the inflammatory edema of the limb (oncometrically), by the content of leukocytes and eosinophils in the blood.

The strength of the skin scar in rats was evaluated by two indicators - the force that causes the scar to break, and the relative elongation at break, which characterizes the elasticity of the tissue.

The effect of drugs on wound healing was studied on a model of a flat-bone skin wound.

Pathological and histochemical studies of the joints were performed randomly.

The material (synovial membrane, articular cartilage of the knee joint) was fixed, embedded in paraffin, stained with hematoxylin-eosin, total GAGs were determined with safranin-T, for separate detection of cholesterol, alcian blue staining in acetate buffer (pH-5.8) with different MgCl content was used. ₂ (0.4 M MgCl ₂ when reacting to cholesterol).

The GAG content was assessed visually in points. Thus, the absence of staining with safranin was considered as subthreshold (minimum) for this dye (-). The content of GAGs is focal, light red staining is like a low content of GAGs (+), diffuse light red is like moderate (++), diffuse bright red is pronounced (+++). No alcian blue staining (-), pale blue staining as low (+), blue as moderate (++), bright blue as significant (+++).

Cartilage tissue destruction was caused by 1% papain solution by intravenous administration of 6 injections with an interval of 3-4 days.

The studies were carried out according to a single scheme for all three groups.

Subgroup I of each group received only papain.

Subgroup II of each group received, respectively, drug A (for group No. 1), drug B (for group No. 2), drug C (for group No. 3).

- 14 041409

Subgroup III, by analogy, received preparations A1 (for group No. 1), B1 (for group No. 2) and B1 (for group No. 3).

Subgroup IV received saline.

The test preparations (A, B, C) and reference preparations (A1, B1, C1) were administered 0.5 ml a day after the last injection of papain daily for a month.

At the end of the experiment, the rats were killed. The costal cartilages were separated, cleaned of soft tissues, dried with acetone, and the total GAG content was determined in them according to the method of D.V. Kosyagin. [Kosyagin D.V. - Issues of medical chemistry, 1982, 28, 4, p. 68-71].

According to the results of the study, it was found that with the introduction of papain, the content of GAG in the costal cartilage of rats decreases. Normally, it is equal to 14.25±2.7 mg of uronic acid per 1 g of dry tissue.

After the end of the experiment, the following results were obtained by groups.

Group #1:_______________________________________________________________

Subgroup I II III IV GAG content (mg/g) 10.95±2.1 15.38±2.8 13.83±2.1 14.20±2.5

Group #2:

Subgroup I II III IV GAG content (mg/g) 10.34±2.2 16.01±2.8 13.88±2.2 14.23±3.1

Group #3:

Subgroup I II III IV GAG content (mg/g) 11.35±2.6 17.48±2.2 13.91±2.3 14.21±2.5

As a result, the results of the experiment on rats showed that the presence of sodium metal silver sulfate in the preparations of chondoitin provides a more pronounced chondroprotective effect in case of damage to cartilage tissue by papain. Such preparations provide a better accumulation of GAGs in the costal cartilage of rats compared to preparations that do not contain metallic silver in their composition.

Example 10

The effectiveness of chondroitin sulfate preparations.

Studies were conducted on the following drugs in the form of a tablet dosage form:

drug A.

Means for the treatment of arthrological diseases of the following composition, mg:

chondroitin sulfate - 500 mg;

calcium stearate - 9.5;

crospovidone - 2.5;

povidone K30 - 2;

microcrystalline cellulose - 190;

metallic silver - 0.0005;

drug A1.

Means for the treatment of arthrological diseases of the following composition, mg:

chondroitin sulfate - 500;

calcium stearate - 9.5;

crospovidone - 2.5;

povidone K30 - 2;

microcrystalline cellulose - 190.

drug B.

Means for the treatment of arthrological diseases of the following composition, mg:

chondroitin sulfate - 1000;

calcium stearate - 10.5;

crospovidone - 3.5;

povidone K30 - 2.8;

microcrystalline cellulose - 220;

metallic silver - 0.005.

Preparation B1.

Means for the treatment of arthrological diseases of the following composition, mg:

- 15 041409 chondroitin sulfate - 1000;

calcium stearate - 10.5;

crospovidone - 3.5;

povidone K30 - 2.8;

microcrystalline cellulose - 220.

drug B.

Means for the treatment of arthrological diseases of the following composition, mg:

chondroitin sulfate - 1800;

stearate - 11.5;

crospovidone - 4.5;

povidone K30 - 3.2;

microcrystalline cellulose - 260;

metallic silver - 0.0126.

drug B1.

Means for the treatment of arthrological diseases of the following composition:

chondroitin sulfate - 1800;

calcium stearate - 11.5;

crospovidone - 4.5;

povidone K30 - 3.2;

microcrystalline cellulose - 260.

Standard control methods were used.

Experimental batches of the dosage form of the drug were used.

The study of drugs was carried out on 60 white rats of different sexes and an initial weight of 250-300 g.

The rats were divided into three groups of 20 animals each.

Group No. 1 - trials of preparation A and reference preparation A1.

Group No. 2 - trials of preparation B and comparison preparation B1.

Group 3 - trials of preparation B and reference preparation B1.

Each group was divided into four subgroups.

The effect of the drug on the course of the inflammatory process was assessed by the change in the magnitude of the inflammatory edema of the limb (oncometrically), by the content of leukocytes and eosinophils in the blood.

The strength of the skin scar in rats was evaluated by two indicators - the force that causes the scar to break, and the relative elongation at break, which characterizes the elasticity of the tissue.

The effect of drugs on wound healing was studied on a model of a flat-bone skin wound.

Pathological and histochemical studies of the joints were performed randomly.

The material (synovium, articular cartilage of the knee joint) was fixed, embedded in paraffin, stained with hematoxylin-eosin, and total GAGs were determined with safranin-T. for separate detection of cholesterol, alcian blue staining was used on acetate buffer (pH-5.8) with different contents of MgCl ₂ (0.4 M MgCl ₂ in the reaction for cholesterol).

The GAG content was assessed visually in points. Thus, the absence of staining with safranin was considered as subthreshold (minimum) for this dye (-). The content of GAGs is focal, light red staining is like a low content of GAGs (+), diffuse light red is like moderate (++), diffuse bright red is pronounced (+++). No alcian blue staining (-), pale blue staining as low (+), blue as moderate (++), bright blue as significant (+++).

Cartilage tissue destruction was caused by 1% papain solution by intravenous administration of 4 injections with an interval of 3-4 days.

The studies were carried out according to a single scheme for all three groups.

Subgroup I of each group received only papain.

Subgroup II of each group received, respectively, drug A (for group No. 1), drug B (for group No. 2), drug C (for group No. 3).

Subgroup III, by analogy, received preparations A1 (for group No. 1), B1 (for group No. 2) and B1 (for group No. 3).

IV subgroup received physiological saline.

Rats of I, II and III subgroups were intravenously injected with papain at 10 mg/kg (1% solution) - 4 injections with an interval of 3-4 days.

Animals of the IV (control) subgroup received physiological saline at the same time.

The preparations were administered to experimental animals in crushed form together with food 1 time per day one day after the last injection of papain for 30 days in the following doses:

group No. 1 - 3.02 mg each;

group No. 2 - 5.3 mg each;

group number 3 - 8.9 mg each.

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Claims (16)

At the end of the experiment, the rats were killed. The costal cartilages were separated, cleaned of soft tissues, dried with acetone, and the total GAG content was determined in them according to the method of D.V. Kosyagin. [Kosyagin D.V. - Issues of medical chemistry, 1982, 28, 4, p. 68-71]. According to the results of the study, it was found that with the introduction of papain, the content of GAG in the costal cartilage of rats decreases. Normally, it is equal to 14.25±2.7 mg of uronic acid per 1 g of dry tissue. After the end of the experiment, the following results were obtained by groups. Group #1:_____________________________________________________________ Subgroup I II III IV GAG content (mg/g) 11.15±2.6 15.01±2.8 13.92±2.3 14.21±2.5 Group #2: Subgroup I II III IV GAG content (mg/g) 11.18±2.4 16.01±2.8 14.08±2.2 14.21±2.5 Group #3: Subgroup I II III IV GAG content (mg/g) 11.35±2.6 17.25±2.4 13.92±2.3 14.21±2.5 The results of the experiment on rats showed that the presence of metallic silver sodium sulfate in the composition of chondroitin preparations provides a more pronounced chondroprotective effect in case of damage to cartilage tissue by papain, simulating the accumulation of GAG in the costal cartilage of rats better than preparations that do not contain metallic silver in their composition. CLAIM 1. An agent for the treatment of arthrological diseases, including an effective amount of sodium chondroitin sulfate and at least one auxiliary substance or carrier, characterized in that it additionally contains metallic or colloidal silver in a relative amount of 0.1 to 10 mg/kg, calculated on weight chondroitin sulfate sodium. 2. The agent according to claim 1, which contains silver in a relative amount of 0.2 to 8 mg/kg, calculated on the weight of sodium chondroitin sulfate. 3. The agent according to claim 1, which contains silver in a relative amount of 0.3 to 6 mg/kg, calculated on the weight of sodium chondroitin sulfate. 4. The agent according to claim 1, which contains silver in a relative amount of 0.4 to 5 mg/kg, calculated on the weight of sodium chondroitin sulfate. 5. The agent according to claim 1, which contains silver in a relative amount of 0.5 to 4 mg/kg, calculated on the weight of sodium chondroitin sulfate. 6. The agent according to claim 1, which contains silver in a relative amount of 0.2 to 3 mg/kg, calculated on the weight of sodium chondroitin sulfate. 7. The agent according to claim 1, which contains silver in a relative amount of 0.15 to 2 mg/kg, calculated on the weight of sodium chondroitin sulfate. 8. The agent according to claim 1, which contains silver in a relative amount of 7 to 10 mg/kg, calculated on the weight of sodium chondroitin sulfate. 9. The agent according to claim 1, which contains silver in a relative amount of 4 to 8 mg/kg, calculated on the weight of sodium chondroitin sulfate. 10. The agent according to claims 1 to 9, wherein the colloidal silver is AgBion-2 colloidal nanosilver concentrate. 11. The agent according to claims 1 to 9, wherein the colloidal silver is a KND-S colloidal silver concentrate. 12. An agent according to claims 1 to 9, in which the amount of chondroitin sodium sulfate is from 50 to 2500 mg. 13. An agent according to claims 1 to 9, wherein the amount of chondroitin sodium sulfate is from 100 to 2000 mg. 14. An agent according to claims 1 to 9, in which the amount of chondroitin sodium sulfate is from 300 to 1500 mg. 15. An agent according to claims 1 to 9, in which the amount of chondroitin sodium sulfate is from 500 to 1200 mg. 16. The agent according to claims 1 to 9, in which the amount of chondroitin sodium sulfate is from 50 to -