RU2812547C1 - Germanium compounds with polyols and polybasic carboxylic acids, method of their preparation and use - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to complex germanium compounds containing an acyclic polyol and a polybasic carboxylic acid or a pharmaceutically acceptable salt thereof, as well as optionally a basic amino acid, of general formula (I): Ge[L] _a [С] _b [A] _c (I), where L is an acyclic polyol of the general formula (HOCH₂[CH(OH)]_nCH₂OH), where n = 2, 3 or 4; C is a polycarboxylic acid selected from the group consisting of ethylenediaminetetraacetic acid and succinic acid, or a pharmaceutically acceptable salt thereof; A is basic amino acid; a is an integer from 1 to 4 inclusive, b is an integer from 1 to 2 inclusive, c is an integer from 0 to 3 inclusive, and the sum of a + b + c is less than or equal to 6. The following is also proposed: a method of producing a complex germanium compound, a composition for dental and oral hygiene or for the prevention and/or treatment of diseases of the teeth and oral cavity, containing an effective amount of a germanium complex compound and at least one acceptable target additive or auxiliary substance, and use of the complex compound germanium for the manufacture of products for dental and oral hygiene or for the prevention and/or treatment of diseases of the teeth and oral cavity. The invention makes it possible to obtain complex compounds of germanium, including an acyclic polyol, a polybasic carboxylic acid or a pharmaceutically acceptable salt thereof, and optionally a basic amino acid, with various combinations of these components.

EFFECT: important advantage of the proposed complex compounds when used for dental and oral hygiene, as well as for the prevention and/or treatment of various diseases of the teeth and oral cavity.

14 cl, 8 dwg, 1 tbl, 6 ex

Description

Field of technology to which the invention relates

The invention relates to the field of organoelement chemistry, namely, to complex compounds of germanium with polyhydric alcohols (acyclic polyols) and polybasic carboxylic acids, as well as additionally, if necessary, with basic amino acids.

More specifically, the invention relates to germanium complex compounds with acyclic polyols of the general formula (HOCH ₂ [CH(OH)] _n CH ₂ OH), where n=2, 3 or 4, which also contain a polycarboxylic acid selected from the group consisting of succinic acid and ethylenediaminetetraacetic acid, or a pharmaceutically acceptable salt thereof, and optionally a basic amino acid.

The resulting complex compounds of germanium with acyclic polyols and polybasic carboxylic acids, optionally containing a basic amino acid, can be used for dental and oral hygiene and/or for the prevention and/or treatment of various diseases of the teeth and oral cavity, such as gingivitis, periodontitis, caries, as well as in areas such as cosmetics and medicine.

State of the art

Acyclic polyols, such as xylitol, sorbitol, erythritol and others, are widely used in the preparation of toothpastes and various pharmaceuticals.

It is known that ordinary dietary carbohydrates affect the precipitation of calcium salts in areas of mineral deficiency in enamel and dentin. In turn, the presence of polyols such as xylitol, D-sorbitol, mannitol and maltitol slows down the formation of turbidity in saliva and inhibits the formation of sediments even in the presence of dextrans and starch (Mäkinen, KK, Söderling, E., Peacor, DR et al. Carbohydrate -controlled precipitation of apatite with coprecipitation of organic molecules in human saliva: Stabilizing role of polyols. Calcif Tissue Int 44 , 258-68 (1989).

It is widely known that periodontal diseases such as gingivitis and periodontitis, as well as dental caries, are associated with the formation of oral biofilms (plaque) (Pihlstrom, B. L., Michalowicz, B. S. & Johnson, N. W. Periodontal diseases. The Lancet 366, 1809-1820 ( 2005)). Dental caries forms over time as a result of a complex interaction between acid-forming bacteria and fermentable carbohydrate. Sugar alcohols, such as xylitol, are among the most widely used oral hygiene substances due to their ability to inhibit biofilm formation. It has also recently been reported that the use of cationic complexes with coordination bonds between zinc chloride and a polyol (erythritol, xylitol, sorbitol) increases the efficiency of biofilm destruction (Lim, J.H., Y. Jeong, S.H. Song, J.H. Ahn, J.R. Lee, and S.M. Lee. Penetration of an antimicrobial zinc-sugar alcohol complex into Streptococcus mutans biofilms. Scientific Reports 1(8): 16154 (2018).

In recent years, a large number of works have been published on the complexation of metal cations with sugars and polyols. Complexes of polyols with cations Ca(II), Cu(II), Fe(II), Mn(II) and other metals are of interest due to their contribution to various biological processes. Coordination between metal and polyol can occur in many biochemical processes, for example in the remineralization of tooth enamel lesions due to the release of Ca(II).

US Pat. No. 8,153,687 has shown that gallium polyalcohol complexes are useful in pharmaceutical applications, particularly for the treatment of infections and infection-associated biofilms. Polyhydric alcohols used in US 8153687 include arabitol, erythritol, sorbitol, mannitol, xylitol and others.

Previously, the authors of the present invention have already reported a method for preparing germanium complex compounds with acyclic polyols, which additionally contain an amino acid or hydroxy acid (RU 2752934).

For animals and humans, germanium is a biologically active and essential trace element, which is confirmed by its presence in almost all human organs and tissues. Germanium is one of the trace elements involved in metabolic processes in the human body. The recommended daily dose of germanium is 0.4-1.5 mg.

Some of the present inventors' early studies indicate that Ge is an effective immune stimulant, and small doses of it can stimulate the production of IgG and IgA, as well as interferons. As is well known, Ig A is the main immunoglobulin in saliva and the oral cavity, responsible for local immunity.

A modern oral hygiene product must cope with the initial stages of gingivitis without masking advanced pathology, which requires examination and treatment in the dentist’s chair. Failure to promptly seek qualified help can lead to the development of severe periodontitis, which may result in the need for tooth extraction due to mobility.

Thus, in the field of dental and oral hygiene, as well as the prevention and/or treatment of diseases of the teeth and oral cavity, in particular in the field of treatment of periodontal diseases, there is a need for new products that can have a comprehensive beneficial effect on the condition of the teeth and oral cavity, in particular, to cope with diseases of the teeth and oral cavity in the initial stages of the disease without the need to use antiseptics.

Disclosure of the invention

An object of the present invention is to provide novel germanium complexes with acyclic polyols which further comprise a polycarboxylic acid selected from the group consisting of succinic acid and ethylenediaminetetraacetic acid, or a pharmaceutically acceptable salt thereof, and optionally a basic amino acid.

Another object of the invention is to provide a method for preparing germanium complexes with acyclic polyols which further contain a polycarboxylic acid selected from the group consisting of succinic acid and ethylenediaminetetraacetic acid, or a pharmaceutically acceptable salt thereof, and optionally a basic amino acid.

Another object of the present invention is to provide a composition for dental and oral hygiene or for the prevention and/or treatment of various dental and oral diseases.

Another purpose of the invention is the use of the obtained germanium complex compounds for the manufacture of products for dental and oral hygiene or for the prevention and/or treatment of various diseases of the teeth and oral cavity.

Thus, in its first aspect, the present invention relates to a germanium complex compound containing an acyclic polyol and a polycarboxylic acid or a pharmaceutically acceptable salt thereof, and optionally a basic amino acid, of general formula (I):

where L is an acyclic polyol of the general formula (HOCH ₂ [CH(OH)] _n CH ₂ OH),

where n = 2, 3 or 4;

C is a polycarboxylic acid selected from the group consisting of ethylenediaminetetraacetic acid and succinic acid, or a pharmaceutically acceptable salt thereof;

A - basic amino acid;

a - represents an integer from 1 to 4 inclusive,

b - represents an integer from 1 to 2 inclusive,

c - represents an integer from 0 to 3 inclusive,

where the sum a + b + c is less than or equal to 6;

in this case, all L in the complex compound are the same or different.

In one embodiment of the germanium complex, the acyclic polyol [ L ] is selected from the group consisting of erythritol, xylitol, and sorbitol.

In yet another embodiment of the germanium complex, the pharmaceutically acceptable polycarboxylic acid salt [ C ] is a sodium salt. In a preferred embodiment, the ethylenediaminetetraacetic acid salt is a disodium salt.

In a further embodiment of the germanium complex, the basic amino acid is selected from the group consisting of arginine, lysine and histidine. In a preferred embodiment, the basic amino acid is selected from the group consisting of arginine and lysine.

In another aspect, the present invention relates to a method for producing the above-described germanium complex compound, comprising the steps of preparing an aqueous suspension of germanium dioxide, adding an acyclic polyol solution to the resulting aqueous suspension of germanium dioxide, heating and stirring the resulting mixture at a temperature of 80-95°C in for 1-5 hours, add a polybasic carboxylic acid or its pharmaceutically acceptable salt, as well as optionally a basic amino acid, to the resulting clear aqueous solution, stir the resulting mixture at a temperature of 60-80°C for 1-3 hours, filter and concentrate the resulting solution using the method partial removal of water, the complex compound is precipitated by adding ethyl alcohol, the alcohol solution is decanted or filtered, and the complex compound is dried by heating under vacuum.

In a further aspect, the present invention relates to a composition for dental and oral hygiene or for the prevention and/or treatment of dental and oral diseases, comprising an effective amount of a germanium complex compound described above and at least one suitable additive or excipient.

In one embodiment, the composition is a dental/oral rinse.

In yet another embodiment of the composition, dental and oral disease is selected from periodontal disease and dental caries. In a preferred embodiment of the composition, the periodontal disease is selected from gingivitis and periodontitis.

Finally, in another aspect, the present invention relates to the use of a germanium complex compound described above for the manufacture of a dental and oral hygiene product or for the prevention and/or treatment of dental and oral diseases.

In one embodiment of the use of a germanium complex compound, dental and oral disease is selected from periodontal disease and dental caries. In a preferred embodiment, the periodontal disease is selected from gingivitis and periodontitis.

Thus, the present invention is directed, in particular, to expanding the arsenal of available germanium complex compounds with acyclic polyols through compounds that additionally include polybasic carboxylic acids selected from the group consisting of succinic acid and ethylenediaminetetraacetic acid, or their pharmaceutically acceptable salts, and optionally basic amino acids. The germanium complex compound of the present invention has both prophylactic and differential diagnostic functions. A polyol organogermanium complex can control initial inflammation and provide symptomatic improvement, which can delay a visit to the dentist without the threat of losing a healthy tooth. The use of the germanium complex compound of the present invention in the early stages of the development of the disease makes it possible to avoid the use of antiseptics, such as chlorhexidine, triclosan, etc.

Possible indications for the use of polyol-organogermanium complex in the oral cavity are, in particular:

- bleeding;

- inflammation of the gingival margin;

- gingivitis;

- stage 1 periodontitis without an autoimmune component;

- acute oral injuries;

- herpetic stomatitis;

- poor oral hygiene;

- removable and non-removable structures in the oral cavity that interfere with high-quality oral hygiene;

- increasing the barrier function of the oral epithelium;

- improvement of colonization resistance of the oral mucosa;

- dental caries.

The present invention allows the composition of the germanium complex to be adjusted and various combinations to be prepared that include a polyol, a polycarboxylic acid or a pharmaceutically acceptable salt thereof, and optionally a basic amino acid. This is an important advantage of the proposed complex compounds when used for dental and oral hygiene or for the prevention and/or treatment of various diseases of the teeth and oral cavity, in particular, such as gingivitis and periodontitis, as well as dental caries, since it makes it possible to obtain compounds with increased antibacterial activity.

The claimed method provides flexibility in the preparation of complex germanium compounds, including a polyol, a polycarboxylic acid or a pharmaceutically acceptable salt thereof, and optionally a basic amino acid, with various combinations of these components depending on the needs and purposes of their subsequent use.

Brief description of drawings

Fig. 1 - ¹ H NMR spectrum in D ₂ O of a complex compound of germanium with xylitol and disodium salt of ethylenediaminetetraacetic acid.

Fig. 2 - ¹ H NMR spectrum in D ₂ O of a complex compound of germanium with sorbitol and disodium salt of ethylenediaminetetraacetic acid.

Fig. 3 - ¹ H NMR spectrum in D ₂ O of a complex compound of germanium with xylitol, succinic acid and arginine.

Fig. 4 - ¹ H NMR spectrum in D ₂ O of a complex compound of germanium with erythritol, succinic acid and arginine.

Fig. 5 - Micrograph of buccal epithelial cells after cultivation for 90 minutes in the presence of a germanium complex according to the invention of GY (A) and chlorhexidine CG (B). Uv. x40.

Fig. 6 - Subjective sensations of patients with gingivitis and periodontitis of the 1st severity before and after a 7-day course of rinsing with a germanium complex according to the invention of GY and chlorhexidine CG and objective data of clinical hygienic indices.

Fig. 7 - Cytological picture in patients with gingivitis and periodontitis of the 1st degree of severity before and after a 7-day course of rinsing with GY and CHG. EB - basophilic epithelium, EE - eosinophilic epithelium, EMKR - punctate microorganisms in/on the epithelium, MKRnaE - multiple microorganisms on the epithelial membrane, MKR - microorganisms in the field of view on glass, GR - granulocytes (neutrophils and macrophages), Gra - activated granulocytes, GRE - granulocytes on the epithelium, GRMKR - granulocytes on microorganisms on glass, GRS - quiet granulocytes, L - lymphocytes, Er - erythrocytes.

Fig. 8 - Colonization function of the buccal epithelium in a clinical experiment before and after rinsing with the germanium complex according to the invention of GY (A and B, respectively) or chlorhexidine CG (C and D, respectively). Uv. x60.

Carrying out the invention

The present invention relates to germanium complex compounds containing acyclic polyols (also known as sugar alcohols), which additionally contain a polycarboxylic acid or a salt thereof, and optionally additionally contain a basic amino acid, and the composition of which is described by the following structural formula:

where L is an acyclic polyol of the general formula (HOCH ₂ [CH(OH)] _n CH ₂ OH),

where n = 2, 3 or 4;

C is a polycarboxylic acid selected from the group consisting of succinic acid and ethylenediaminetetraacetic acid, or a pharmaceutically acceptable salt thereof;

A - basic amino acid;

a - represents an integer from 1 to 4 inclusive,

b - represents an integer from 1 to 2 inclusive,

c - represents an integer from 0 to 3 inclusive,

in this case, all L in the complex compound are the same or different.

In structural formula (I), a takes values from 1 to 4, more preferably 2 or 3; b takes the value 1 or 2, more preferably the value 1; c takes values from 0 to 3, more preferably the value 2. In this case, the sum a+b+c is less than or equal to 6.

As germanium dioxide, germanium dioxide with a hexagonal crystal structure can be used, which is insoluble in water and, when mixed with water, forms a suspension in water.

As acyclic polyols with a chain length of 4 (n=2), 5 (n=3) or 6 (n=4) carbon atoms, various stereoisomers of butane-1,2,3,4-tetrol can be used, respectively, such as , for example, erythritol or threitol; various stereoisomers of pentane-1,2,3,4,5-pentol, such as, for example, xylitol, ribitol or arabitol; various stereoisomers of hexane-1,2,3,4,5,6-hexol, such as, for example, mannitol, iditol, galactitol or sorbitol. For the purposes of the present invention, acyclic polyols such as erythritol, xylitol and sorbitol are preferred.

For the purposes of the present invention, preferred polycarboxylic acids are ethylenediaminetetraacetic acid (EDTA) or its pharmaceutically acceptable salts, for example, disodium salt (also known as complexone III or Trilon B), and succinic acid or its salts. Preferred salts of these polybasic acids are sodium salts.

As is known to one skilled in the art, essential amino acids include the amino acids lysine, arginine and histidine. All of these amino acids are capable of carrying a positive charge because the functional groups present in the side chains of the said amino acids can accept a proton. The lysine side chain functional group is an amino group, the arginine side chain functional group is a guanidine group, and the histidine side chain functional group is an imidazole group. The basis of this ability, therefore, is the presence in the functional group of the side chain of a nitrogen atom, which carries a lone electron pair and can serve as its donor for interaction with a proton. This ability to accept a proton determines the similarity of lysine, arginine and histidine residues with respect to functional activity. Thus, in the composition of proteins, arginine, lysine and histidine residues are often interchangeable, so the replacement of one of these amino acid residues with another is considered to be conservative. For the purposes of the present invention, arginine and lysine are preferred as basic amino acids.

These amino acids, taking into account their zwitterionic nature, can be presented both in the form of a free acid/base and in a salt form.

In the context of the present invention, the expression "pharmaceutically acceptable" refers in particular to acid addition salts formed by pharmaceutically acceptable inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like; or pharmaceutically acceptable organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo[ 2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tert- butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, p-toluenesulfonic acid, etc.

In the context of the present invention, the expression "pharmaceutically acceptable" also refers to base addition salts formed by pharmaceutically acceptable organic bases such as ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, ammonia, cyclohexylamine, dicyclohexylamine and the like; or pharmaceutically acceptable inorganic bases such as aluminum hydroxide, calcium hydroxide, magnesium hydroxide, potassium carbonate, potassium hydroxide, sodium carbonate, sodium hydroxide and the like.

The method according to the invention is characterized by the fact that an aqueous solution of an acyclic polyol is added to an aqueous suspension of germanium dioxide with stirring. According to the method, one acyclic polyol or a mixture of several different acyclic polyols can be added. The mixture is stirred at 80-95°C for 1-5 hours until a clear solution is obtained. Next, a polybasic carboxylic acid or a pharmaceutically acceptable salt thereof, as well as optionally a basic amino acid, is added to the resulting solution, and the mixture is stirred at 60-80°C for 1-3 hours until a clear solution of the target product is obtained. Then the solution is filtered and concentrated by distilling off part of the water and the product is precipitated by adding ethyl alcohol.

To isolate complex compounds of germanium with polyols and polybasic carboxylic acids or their pharmaceutically acceptable salts, optionally containing a basic amino acid, the precipitate is separated from the solution by filtration or decantation and then dried on a rotary evaporator to remove residual water and alcohol.

A composition for dental and oral hygiene or for the prevention and/or treatment of dental and oral diseases of the present invention, containing a complex compound of germanium with a polyol and a polybasic carboxylic acid or a pharmaceutically acceptable salt thereof, optionally containing a basic amino acid, may be presented in various forms, for example, in the form of toothpaste, dentifrice, tooth powder, pre-brushing mouth rinse, post-brushing mouth rinse, topical oral gel, chewing gum, lozenges, or any other form suitable for use on the surfaces of the mouth, teeth or gums.

The composition for dental and oral hygiene and/or for the prevention/treatment of dental and oral diseases of the present invention contains a germanium complex compound with a polyol and a polybasic carboxylic acid or a pharmaceutically acceptable salt thereof, optionally containing a basic amino acid, in an effective amount. In the context of the present invention, the expression "effective amount" means an amount that is sufficient to achieve the desired hygienic or prophylactic/therapeutic effect in the person using the composition, without causing excessive harmful side effects (such as toxicity, inflammation, allergic reaction, etc.). ), commensurate with an acceptable benefit/risk ratio. The specific effective amount of the germanium complex compound of the present invention will vary depending on factors such as the particular condition being cared for or prevented/treated, the physical condition of the subject, the particular germanium complex compound used, the particular form in which the composition of the present invention is presented, desired application scheme, etc.

In addition to the complex compound of germanium with a polyol and a polycarboxylic acid or a pharmaceutically acceptable salt thereof, optionally containing a basic amino acid, the compositions of the present invention may contain standard ingredients, various targeted additives and excipients traditionally used in compositions for the specified purpose, depending on the form of their implementation and are acceptable for use in the oral cavity. Thus, without limitation, toothpastes or gels may contain an abrasive material, a moisture-retaining substance, a cleansing and foaming agent, in particular, a surfactant, a source of fluoride, a preservative, a sweetener, a fragrance, a formative thickener, a binder, and various biologically active additives, water, colorant, chelating agent, adhesion promoter, pH adjusting agent, colorant, flavoring agent, flavoring agents, other targeted additives, etc.

Without limitation, mouth rinses may contain an aqueous medium, a moisture-retaining additive, a preservative, a dye, a flavoring agent, a pH regulator, a formative thickener, a solubilizer, a film former, a biologically active additive, a flavoring agent, a source of fluoride, other targeted additives, etc.

Specific substances traditionally used in the manufacture of hygienic and prophylactic/therapeutic pastes and gels, as well as various rinses for teeth and mouth, are well known to specialists in the art.

For example, silicon dioxide, calcium carbonate (chalk), titanium dioxide, etc. can be used as an abrasive material.

Glycerin, sorbitol, etc. can be used as a humectant.

Sodium lauryl sulfate (sodium dodecyl sulfate), sodium lauroyl sarcosinate, sodium N-cocoyl-N-methyl taurate, etc. can be used as a surfactant.

Sodium fluoride, potassium fluoride, sodium monofluorophosphate, copper fluoride, tin fluoride, ammonium fluorosilicate, sodium fluorozirconate, ammonium fluorozirconate, aluminum fluorophosphates (mono-, di- and tri) can be used as a source of fluorine; fluorinated sodium calcium pyrophosphate, etc., as well as mixtures thereof.

As a sweetener, for example, sodium saccharinate, aspartame, cyclamate, sucralose, honey herb preparations (stevia), mannitol, sorbitol, xylitol and similar polyols, etc. can be used.

Monosodium salt of para-hydroxybenzoic acid methyl ester or sodium salt of para-hydroxybenzoic acid propyl ester, etc. can be used as a preservative.

Compounds of both plant origin and chemically synthesized compounds, including various plant extracts, etc. can be used as biologically active substances.

The formative thickener may be selected from natural or synthetic agents including carrageenan, hydroxymethylcellulose, silica or colloidal silica thickeners, methyl vinyl ether-maleic anhydride, sodium carboxymethylcellulose, etc.

Methods for preparing hygienic or therapeutic compositions of the present invention containing a complex compound of germanium with a polyol and a polycarboxylic acid or a salt thereof, optionally containing an amino acid or a salt thereof, are themselves well known to those skilled in the art.

Below, preferred embodiments of the claimed inventions are illustrated by examples of implementation, which, however, are not intended to limit the scope of the claims, which is determined solely by the attached claims.

Example 1. Preparation of a complex compound of germanium with xylitol and disodium salt of ethylenediaminetetraacetic acid.

250 ml of distilled water and 2.1 g of germanium dioxide, GeO ₂ are loaded into a 4-neck round-bottomed flask with a volume of 0.5 l, equipped with a stirrer and a thermometer. 100 ml of an aqueous solution of polyol containing 12.2 g of xylitol, HO(CH ₂ )(CHOH) ₃ (CH ₂ )OH, is added to an aqueous suspension of germanium dioxide with stirring and heating. The suspension is stirred at 90-95°C for 2 hours until a clear solution is formed. Next, 7.1 g of disodium salt of ethylenediaminetetraacetic acid, (HOOCCH ₂ ) ₂ N(CH ₂ ) ₂ N(CH ₂ COONa) ₂ is added to the solution and stirred for 1 hour at 60-70°C. The resulting aqueous solution is cooled to room temperature, filtered and concentrated, removing most of the water on a rotary evaporator. The product is precipitated from the aqueous concentrate by adding ethyl alcohol in a ratio of 1:5 by volume. The precipitate is isolated and dried on a rotary evaporator to remove residual water and alcohol. 16.9 g of powder of a complex compound of germanium with xylitol and disodium salt of ethylenediaminetetraacetic acid, Ge[C ₅ H ₁₂ O ₅ ] ₄ [C ₁₀ H ₁₂ N ₂ Na ₂ O ₈ ], are obtained. The compound is highly soluble in water; a 5% solution has a pH of 5.5. The NMR spectrum of the resulting compound is shown in Fig. 1.

Elemental analysis:

calc.: C 35.47 H 5.91 N 2.76 Na 4.53 Ge 7.15

detect: C 35.39 H 5.99 N 2.81 Na 4.61 Ge 7.08

Example 2. Study of a complex compound of germanium with sorbitol and disodium salt of ethylenediaminetetraacetic acid.

250 ml of distilled water and 2.1 g of germanium dioxide, GeO ₂ are loaded into a 4-neck round-bottomed flask with a volume of 0.5 l, equipped with a stirrer and a thermometer. With stirring and heating, 100 ml of an aqueous solution of a polyol containing 7.3 g of sorbitol, HO(CH ₂ )(CHOH) ₄ (CH ₂ )OH, is added to an aqueous suspension of germanium dioxide. The suspension is stirred at 90-95°C for 2 hours until a clear solution is formed. Next, 6.8 g of disodium salt of ethylenediaminetetraacetic acid, (HOOCCH ₂ ) ₂ N(CH ₂ ) ₂ N(CH ₂ COONa) ₂ is added to the solution and stirred for 1 hour at 60-70°C. The resulting aqueous solution is cooled to room temperature, filtered and concentrated, removing most of the water on a rotary evaporator. The product is precipitated from the aqueous concentrate by adding ethyl alcohol in a ratio of 1:5 by volume. The precipitate is isolated and dried on a rotary evaporator to remove residual water and alcohol. 13.5 g of powder of a complex compound of germanium with sorbitol and disodium salt of ethylenediaminetetraacetic acid, Ge[C ₆ H ₁₄ O ₆ ] ₂ [C ₁₀ H ₁₂ N ₂ Na ₂ O ₈ ], are obtained. The compound is highly soluble in water; a 5% solution has a pH of 5.4. The NMR spectrum of the resulting compound is shown in Fig. 2.

Elemental analysis:

calc.: C 34.26 H 5.24 N 3.63 Na 5.96 Ge 9.41

detect.: C 34.19 H 5.32 N 3.69 Na 6.03 Ge 9.35

Example 3. Preparation of a complex compound of germanium with xylitol, succinic acid and arginine.

250 ml of distilled water and 2.1 g of germanium dioxide, GeO ₂ are loaded into a 4-neck round-bottomed flask with a volume of 0.5 l, equipped with a stirrer and a thermometer. With stirring and heating, 100 ml of an aqueous solution of a polyol containing 9.2 g of xylitol, HO(CH ₂ )(CHOH) ₃ (CH ₂ )OH, is added to an aqueous suspension of germanium dioxide. The suspension is stirred at 90-95°C for 2 hours until a clear solution is formed. Next, 2.4 g of succinic acid and HOOC(CH ₂ ) ₂ COOH are added to the solution and stirred for 1 hour at 60-70°C. Then an additional 7.0 g of arginine, HN=C(NH ₂ )NH(CH ₂ ) ₃ CH(NH ₂ )COOH, is added and stirred for 1 hour at 60-70°C. The resulting aqueous solution is cooled to room temperature, filtered and concentrated, removing most of the water on a rotary evaporator. The product is precipitated from the aqueous concentrate by adding ethyl alcohol in a ratio of 1:5 by volume. The precipitate is isolated and dried on a rotary evaporator to remove residual water and alcohol. 18.1 g of powder of a complex compound of germanium with xylitol, succinic acid and arginine, Ge[C ₅ H ₁₂ O ₅ ] ₃ [C ₄ H ₆ O ₄ ][C ₆ H ₁₄ N ₄ O ₂ ] ₂ , are obtained. The compound is highly soluble in water; a 5% solution has a pH of 5.6. The NMR spectrum of the resulting compound is shown in Fig. 3.

Elemental analysis:

calc.: C 37.40 H 7.01 N 11.25 Ge 7.29

detect.: C 37.34 H 7.09 N 11.17 Ge 7.21

Example 4. Preparation of a complex compound of germanium with erythritol, succinic acid and arginine.

250 ml of distilled water and 2.1 g of germanium dioxide, GeO ₂ are loaded into a 4-neck round-bottomed flask with a volume of 0.5 l, equipped with a stirrer and a thermometer. With stirring and heating, 100 ml of an aqueous solution of a polyol containing 9.8 g of erythritol, HO(CH ₂ )(CHOH) ₂ (CH ₂ )OH, is added to an aqueous suspension of germanium dioxide. The suspension is stirred at 90-95°C for 2 hours until a clear solution is formed. Next, 2.4 g of succinic acid and HOOC(CH ₂ ) ₂ COOH are added to the solution and stirred for 1 hour at 60-70°C. Then an additional 3.5 g of arginine, HN=C(NH ₂ )NH(CH ₂ ) ₃ CH(NH ₂ )COOH, is added and stirred for 1 hour at 60-70°C. The resulting aqueous solution is cooled to room temperature, filtered and concentrated, removing most of the water on a rotary evaporator. The product is precipitated from the aqueous concentrate by adding ethyl alcohol in a ratio of 1:5 by volume. The precipitate is isolated and dried on a rotary evaporator to remove residual water and alcohol. 15.4 g of powder of a complex compound of germanium with erythritol, succinic acid and arginine, Ge[C ₄ H ₁₀ O ₄ ] ₄ [C ₄ H ₆ O ₄ ][C ₆ H ₁₄ N ₄ O ₂ ], are obtained. The compound is highly soluble in water; a 5% solution has a pH of 4.7. The NMR spectrum of the resulting compound is shown in Fig. 4.

Elemental analysis:

calc.: C 36.59 H 7.03 N 6.56 Ge 8.51

detect.: C 36.63 H 7.08 N 6.51 Ge 8.45

Example 5. Preliminary study of the effectiveness and toxicity of a polyol organogermanium complex.

To preliminary assess the effectiveness and toxicity of the complex, an in vitro culture test of the buccal epithelium of 5 healthy volunteers with intact periodontal tissues was conducted. The subjects did not brush their teeth or eat for 4 hours before collecting the epithelium with a microbrush. Cultivation of the buccal epithelium was carried out for 90 minutes at a temperature of 37°C in the presence of the test complex described in Example 3 at a concentration of 5% in saline solution (HS) or a control reference antiseptic - chlorhexidine bigluconate at a concentration of 0.02% (CH). Epithelium sampling and cultivation were carried out daily for 7 days to obtain a sufficient number of samples (30 smears in each of the HY and CG treatment groups). Smears taken from each patient were transferred to 2 glasses for subsequent parallel cultivation in the presence of GY or CG. After cultivation, the smears were left to air dry, fixed with 95% ethyl alcohol, and stained with hematoxylin-eosin.

Microphotographs of buccal epithelial cells after cultivation for 90 minutes in the presence of GY or CG are presented in Fig. 5. In the HY treatment group (Fig. 5A), basophilic coloring predominates, a minimal number of microorganisms (Mkr) on the glass in the field of view and a moderate number of microorganisms on epithelial cells. Activated immunocytes in moderate quantities. The epithelium is mature, ready for desquamation. There were no signs of toxigenic granularity or cytoplasmic eosinophilia. Based on the microscopy results, it is clear that samples of buccal epithelium cultured in the presence of GN had a uniform tendency towards activation of the barrier and colonization functions of the epithelium, moderate activation of granulocytes, pronounced inhibition of microbial adhesion to glass, but good adhesion to the membrane of mature epithelial cells (normalization of colonization function - the zero line of defense of the innate immunity of the mucous membranes, while the activation of granulocytes is the first). It is important to note the absence of a predominance of eosinophilic staining of the cytoplasm of epithelial cells, which indicates the absence of an inflammatory component and the negative/toxic effect of OH on the mucosal epithelium.

In the hCG treatment group (Fig. 5B), the epithelium has a sharply eosinophilic color, multiple microorganisms are observed on the glass and on epithelial cells, and multiple activated immunocytes. The entire field of view is covered with eosinophilic chlorhexidine crystals. After culturing the buccal epithelium of healthy volunteers in the presence of CG, a homogeneous picture was observed from all glasses: bright eosinophilic coloring of epithelial cells or rarely - eosinophilic-basophilic, multiple crystallization of CG on the glass, multiple MCRs (it is possible that dead ones that are not washed off from the glass are so as recorded by hCG crystals), granulocytes are sharply activated with membrane ruptures, lymphocytes are activated (single), colonization resistance of epithelial cells is sharply reduced or absent.

It is noteworthy that after cultivation in a thermostat (15 minutes), chlorhexidine evaporated and crystallized many times faster in open air compared to the organogermanium complex, which essentially did not evaporate or crystallize for 10 hours. The GY solution had an excellent “washing away” effect against microorganisms in the smear. The high content of microorganisms in the smear of the CG group can be explained by the high degree of crystallization of chlorhexidine on the glass and thereby “sealing” the entire contents of the smear without washability and self-cleaning.

Table 1 presents the parameters of the cytological portrait after culturing the buccal epithelium in the presence of GY and CH.

Table 1. Cytological portrait after culturing the buccal epithelium for 90 minutes in the presence of GY and CH Morphological element in the field of view GY group
(average spread in one field of view) HG Group
(average spread in one field of view) Functional significance Epithelium basophilic 13-47 6-27 Calm healthy epithelium Epithelium eosinophilic 2-8 6-18 Activated or inflamed Epithelial cells with microns on the membrane 3-9 14-30 Normal barrier colonization function Microdistrict single on the epitome or in it 0-11 15-30 Colonization barrier function is impaired Microdistrict on glass 12-37 Multiple more than 30% of the field of view 200-500 Risk of plaque and contamination/progression of the infectious process Granulocytes in the field of view 2-7 7-25 The norm is if not more than 10-15 Activated granulocytes 2-7 7-25 Fighting infection Granulocytes on the epithelium 1-3 4-23 Risk of auto-aggression Granulocytes on microdistrict 0-2 2-5 Normal first line immune defense Granulocytes are “quiet” 0 0 There is no risk of auto-aggression and there is good protection against infection Lymphocytes 1-5 1-3 Antiviral or autoimmune component % deviation from glass to glass No more than 30% No more than 18% The result is reproducible and reliable at a rate of less than 30% % deviation from visual field to visual field No more than 50% No more than 20% The result is reproducible and reliable when the rate is less than 50%

Example 6. Study of the effect of an organogermanium complex on bleeding, the inflammatory component and epithelial-immune interactions of the gums in patients with gingivitis/periodontitis of the 1st severity.

A clinical pilot study of the effectiveness of the organogermanium complex was carried out on a group of patients with signs of inflammation of marginal periodontal tissue (with gingivitis/periodontitis of the 1st severity). The patients were divided into 2 groups: the first group (n=3) used the organogermanium complex described in Example 3 as a rinse (in the form of a 5% solution in saline) in the morning and evening after brushing their teeth for 7 days, the second group (n=3 ) for the same purposes used a solution of chlorhexidine bigluconate at a concentration of 0.02%. The patients' condition was assessed before and after rinsing teeth for 7 days with the studied compositions based on a diagnostic complex, which included:

1. A scale for subjective assessment of the condition of the oral tissues by the patient: tooth sensitivity, plaque, bleeding. The evaluation criterion is self-feeling on a scale from 1 to 5, where 1 is the minimum and 5 is the maximum;

2. Coloring the degree of glycogen accumulation in the epithelium of the marginal periodontium (Schiller-Pisarev Test), as a sign of the inflammatory process;

3. Plaque staining in patients according to Fedorova-Volodkina;

4. Gingivitis index PMA + Schiller-Pisarev;

5. PBI bleeding index;

6. Light microscopy with hematoxylin-eosin staining of cytological smears.

To study the colonization function of the buccal epithelium, additional sampling of the buccal epithelium was carried out in a clinical experiment before and after rinsing with HY and CHG in patients.

Simple statistical processing was carried out with comparative graphical visualization of the results.

The results of the patient's subjective assessment of the condition of the oral tissues before and after a 7-day course of rinsing with HY and the reference drug CG are presented in Fig. 6. After rinsing with GY, patients noted a decrease in gum bleeding, tooth sensitivity, plaque and a feeling of “inflammation”. After rinsing with hCG, patients noted a decrease in gum bleeding, increased tooth sensitivity, plaque, and a feeling of “inflammation.” Interestingly, all patients in the GY group noticed a sharp cessation of bleeding on the 5th day, while patients in the CG group either did not notice a decrease in bleeding or noted a slight gradual decrease from the first to the seventh day.

The clinical index picture (Fig. 6) showed a decrease in the hygiene index, a significant decrease in the bleeding and gingivitis indices after rinsing the GJ. After a 7-day rinse with hCG, there was a decrease in the bleeding index, while the Fedorova-Volotkina hygiene and gingivitis indices were slightly increased.

The results of the analysis of the cytological picture in patients before and after a 7-day course of rinsing with HY and CH are presented in Fig. 7. In the cytological smears of the GY treatment group on the 7th day, a total increase in the epithelium was observed by 90%, an increase in the epithelium carrying microorganisms by 97%, an increase in the number of granulocytes on the epithelium (with microorganisms) by 151% and a decrease in the number of erythrocytes in smear by 100%, reducing the number of microorganisms in the smear by 41%.

In the cytological smear of the hCG treatment group (Fig. 7) on the 7th day, a decrease in the number of epithelium with microorganisms on the epithelial membrane was found by 69%, a decrease in the number of microorganisms in the smear by 76%, an increase in the number of granulocytes by 397%, quiet, non-activated granulocytes increased in number in relation to the data at the beginning of the experiment by 16 times, 4.5 times more granulocytes settled on the microorganisms of the smear outside the epithelial cells than before the start of the hCG rinsing course, the number of lymphocytes increased by 263%. The number of red blood cells in smears decreased by 33%.

In FIG. Figure 8 shows the results of a study of the colonization barrier function of the buccal epithelium in patients before and after rinsing with HY and CH. In the group of GY treatment before rinsing (Fig. 8A), even in the presence of an inflammatory component (eosinophilic cytoplasm, single microorganisms on the epithelial cell membrane, no clear contours of the membrane), on the 7th day after rinsing, basophilic-eosinophilic staining of the cytoplasm and multiple microorganisms are noted on the membrane of epithelial cells (Fig. 8B).

In the CG treatment group, the opposite picture is observed - a decrease in the basophilicity and colonization efficiency of epithelial cells after rinsing (Fig. 8D) even under the starting conditions of high colonization efficiency of the buccal epithelium (Fig. 8C).

The study demonstrates the high efficiency of the developed polyol organogermanium complex for the correction of the initial stages of periodontal tissue disease (gingivitis and periodontitis of the 1st degree). The chlorhexidine bigluconate solution used as a comparison sample turned out to be ineffective in the case of early manifestations of gingivitis and periodontitis of the 1st degree of severity. Thus, the use of the polyol organogermanium complex of the present invention makes it possible to avoid the use of antiseptics such as chlorhexidine, triclosan, etc. in the early stages of disease development.

In addition to objective clinical data, attention should be paid to the patient’s self-feelings upon first contact with a hygiene product, which influences the patient’s further preferences in purchase and regularity of use. In the experiment, patients often noted the appearance of increased tooth sensitivity and plaque after rinsing with chlorhexidine, which caused a feeling of “increasing inflammation in the oral cavity.” Objective data confirms the subjective assessment of patients, since chlorhexidine was strongly eosinophilic (acidic pH value) in the culture test and showed a pronounced tendency to crystallize, had a reduced “washing” ability, which could be the cause of the sensations described above. In the GY treatment group, patients did not feel any visible effect until the fifth day. On the 5th day, they noticed that “the bleeding immediately disappeared.” This phenomenon can be explained by the pattern of the pathophysiological process of inflammation and the composition of the germanium complex.

It is known that inflammatory phenomena in uncomplicated inflammation increase by the 4th day, and without the addition of a bacterial infection, chronic injury, autoimmune, allergic components, the process subsides, and from the 5-7th day the regeneration process starts (Zaichik A.Sh., Churilov L.P. Fundamentals of pathochemistry (Textbook for university students) - St. Petersburg, 2001. - ELBI-SPb, 688 pp.). The proposed organogermanium complex, including germanium, xylitol, succinic acid and arginine, contributes to the normal resolution of the inflammatory focus.

The effectiveness of the organogermanium polyol complex of the present invention was confirmed in clinical and morphological experiments. In the GY treatment group, an increase in colonization resistance of the epithelium and a decrease in the inflammatory reaction are clearly visible (eosinophilic cytoplasm, single microorganisms on the epithelial cell membrane, no clear contours of the membrane); on the 7th day of rinsing, basophilic-eosinophilic coloring of the cytoplasm and multiple microorganisms on the membrane of the epithelial cell, ready to desquamation - pyknotic nucleus. The increase in desquamation is determined by the total increase in the epithelium, which was 90%, the epithelium carrying microorganisms, by 97%, in the group of treatment with GY with gingivitis and periodontitis of the 1st severity on the 7th day of rinsing. Desquamation and colonization resistance represents the first barrier epithelial line of defense of the oral mucosa. In the event that the epithelial barrier does not work as a self-cleaning process through increased desquamation of epithelial cells with adherent microorganisms, granulocytes are attracted to the site of inflammation (due to pro-inflammatory signals of epithelial cells, in a cytological smear - eosinophilic inclusions of glycogen in the cytoplasm of the epithelium) (Lebedev K.A., Ponyakina I.D. Immunology of pattern recognition receptors: Integral immunology. 2nd edition. - M.: Book house "LIBROKOM", 2013. - 256 pp.)

Stabilization of the vascular wall was unequivocally demonstrated in a clinical experiment, both by the subjective sensation of patients (reduced bleeding), and clinically (decrease in hygiene index, bleeding and gingivitis after rinsing the GJ), as well as morphologically (100% decrease in red blood cells in the smears of patients after rinsing the GJ) . In the group of patients, by the 7th day of rinsing the GY, there was an increase in the number of granulocytes on the epithelium (with microorganisms) by 151% and a decrease in the number of microorganisms in the smear by 41% with a general increase in granulocytes by 75% (in contrast to the CG treatment group, an increase in granulocytes in which amounted to 397%). These results indicate an adequate immune response without excessive autoaggression and with sufficient strength to self-cleanse the inflammation in the GY treatment group.

Claims (31)

1. A complex germanium compound containing an acyclic polyol and a polycarboxylic acid or a pharmaceutically acceptable salt thereof, and optionally a basic amino acid, of general formula (I): where L is an acyclic polyol of the general formula (HOCH ₂ [CH(OH)] _n CH ₂ OH), where n = 2, 3 or 4; C is a polycarboxylic acid selected from the group consisting of ethylenediaminetetraacetic acid and succinic acid, or a pharmaceutically acceptable salt thereof; A – basic amino acid; a – represents an integer from 1 to 4 inclusive, b – represents an integer from 1 to 2 inclusive, с – represents an integer from 0 to 3 inclusive, where the sum a + b + c is less than or equal to 6. 2. The germanium complex compound according to claim 1, in which the acyclic polyol [L] is selected from the group consisting of erythritol, xylitol, sorbitol. 3. Complex compound of germanium according to any one of paragraphs. 1, 2, wherein the pharmaceutically acceptable polycarboxylic acid salt [ C ] is a sodium salt. 4. The germanium complex compound according to claim 3, in which the salt of ethylenediaminetetraacetic acid is a disodium salt. 5. The germanium complex compound according to claim 1, in which the main amino acid is selected from the group consisting of arginine, lysine and histidine. 6. The germanium complex compound according to claim 5, in which the main amino acid is selected from the group consisting of arginine and lysine. 7. A method for producing a germanium complex compound according to any one of claims. 1-6, including stages in which: - obtain an aqueous suspension of germanium dioxide, - add a solution of acyclic polyol to the resulting aqueous suspension of germanium dioxide, - heat and stir the resulting mixture at a temperature of 80-95°C for 1-5 hours, - add to the resulting clear aqueous solution a polybasic carboxylic acid or a pharmaceutically acceptable salt thereof, and optionally a basic amino acid, - stir the resulting mixture at a temperature of 60-80°C for 1-3 hours, - filter and concentrate the resulting solution by partial removal of water, - precipitate the complex compound by adding ethyl alcohol, - decant or filter the alcohol solution and dry the complex compound by heating under vacuum. 8. Composition for dental and oral hygiene or for the prevention and/or treatment of dental and oral diseases, containing an effective amount of a germanium complex compound according to any one of claims. 1-6 and at least one suitable additive or excipient. 9. Composition according to claim 8, which is a rinse for teeth/oral cavity. 10. The composition according to claim 8, where the disease of the teeth and oral cavity is selected from periodontal diseases and dental caries. 11. The composition according to claim 10, wherein the periodontal disease is selected from gingivitis and periodontitis. 12. Use of a germanium complex compound according to any one of paragraphs. 1-6 for the manufacture of products for dental and oral hygiene or for the prevention and/or treatment of diseases of the teeth and oral cavity. 13. Use according to claim 12, wherein the disease of the teeth and oral cavity is selected from periodontal diseases and dental caries. 14. Use according to claim 13, wherein the periodontal disease is selected from gingivitis and periodontitis.