RU2755346C1 - Composition containing epigenome modernization factor, which has reparative, oncoprotective and anti-inflammatory effect - Google Patents

Abstract

FIELD: medicine, biotechnology.

SUBSTANCE: invention relates to medicine and biotechnology and can be used to create a remedy that has a reparative and anti-inflammatory effect. The composition, which has an anti-inflammatory and regenerative effect, includes hyaluronic acid and/or its derivatives in an amount of 0.1 to 10 wt.%, a fraction of mitochondrial RNA in an amount of 0.001 to 10 wt.%, 0.9% NaCl solution or water – up to 100 wt.%.

EFFECT: invention provides pronounced resistance to the effects of hyaluronidase.

4 cl, 3 tbl, 13 ex

Description

Technology area

The present invention relates to medicine and biotechnology and can be used to create an agent that has a reparative and anti-inflammatory effect.

State of the art

The problem of accelerating tissue regeneration after injuries of various types attracts the attention of a wide range of specialists, and many approaches are used to solve it. One of the directions in solving this problem is the creation of hydrogels based on hyaluronic acid containing biologically active components.

From the prior art, gels based on hyaluronic acid for cosmetology and treatment of skin lesions are known, which are used externally to moisturize the skin, improve firmness and elasticity by stimulating the production of elastin and collagen, in addition, hyaluronic acid-based gels for injection are used for cosmetic purposes, for filling or replacing biological tissues (for example, filling wrinkles, remodeling the face contour, increasing lip volume, etc.), as well as for skin rehydration treatment using the biorevitalization method, when injections of a pure hyaluronic acid solution are made intradermally, as well as mesotherapy, when therapeutic or prophylactic additives such as vitamins, amino acids, polynucleotides are added to the hyaluronic acid hydrogel for intradermal injection.

Another application of the hyaluronic acid-based gel is its use as intra-articular implants, where the gel is a synovial fluid prosthesis, reduces inflammation and stimulates tissue regeneration (RF patent No. 2683286, publ. 03/27/2019).

There are also known methods of using hyaluronic acid-based gels for the treatment and prevention of diseases of the mucous membranes of the digestive system (RF patent No. 2679638, publ. 06/08/2018), genitourinary system (RF patent No. 2519010, publ. 06/10/2014), respiratory system and mucous membranes (RF patent No. 2697671, publ. 08/16/2019).

In the described methods of application, the bioresorption property of a biopolymer of hyaluronic acid containing biologically active substances added to it is used. Substances are fixed on polymer chains due to chemical and physical bonds and are released into the surrounding tissues as the biopolymer is resorbed due to the action of the enzyme hyaluronidase, and this process can vary in time depending on the structuring of the biopolymer. In this regard, many efforts have been made to improve the physicochemical stability of hyaluronic acid-based gels in order to increase the in vivo period of action of the gel (that is, the residence time of the gel at the injection site) and thereby increase the duration of therapeutic efficacy (RF patent No. 2613887, publ. . 03/21/2017).

These methods use the main biological properties of hyaluronic acid, namely the ability to retain water in tissues, the effect on the intensity of ion exchange in cells, the rate of transport of various biologically active substances and toxins, the impermeability of the medium for large molecules and cells. When used as a prosthesis of synovial fluid in joints, hyaluronic acid, due to its viscosity, acts as a lubricant for rubbing cartilaginous surfaces, and also reduces the negative effect of external pressure on the surface of the joint. The use of cross-linked hyaluronic acid improves its viscoelastic properties and the bioresorption period in vivo under the action of the hyaluronidase enzyme.

The disadvantages of these methods is the absence of auxiliary components in the composition of the proposed compositions as a factor modulating cell regeneration, proliferation and providing a therapeutic (for example, anti-inflammatory) effect.

Nucleic acid components, in particular polydeoxyribonucleotides, are an important component of cells and are used for various purposes, such as in medicine for treating and improving the surface of wounds by injecting these fractions into the area of wounds or the like, and in cosmetology to improve the situation with wrinkles caused by cellular activity (Shapovalova E.Yu., Boyko T.A., Baranovsky Yu.G., Kolomoets T.A., Vasilenko S.A.Morphological features of tissue regeneration in a model ischemic skin wound under the influence of polynucleotides, Crimean journal experimental and clinical medicine, vol. 7, No. 4, 2017).

The closest analogue of the claimed invention is an injection composition containing a hyaluronic acid derivative and a DNA fraction consisting of polynucleotides (RF patent No. 2697671, publ. 08.16.2019), used for subcutaneous injections and for the treatment of dry eye syndrome. The disadvantage of this invention is that to obtain a pronounced proliferative effect, it is necessary to use DNA polynucleotides in high concentration due to their relatively low proliferative activity and a large amount of non-working ballast, consisting of polynucleotides that carry a trophic function. At the same time, comparative laboratory studies have shown that the ratio required to activate the fission process is from 1 to 100 mg of polynucleotides per 1 ml of hydrogel, which leads to a clouding of the final product, an increase in its cytotoxicity, and an increase in the risk of developing allergic reactions when injected.

The objective of the present invention is to develop an agent based on hyaluronic acid having good viscoelasticity, enzymatic stability, with the addition of a fraction of mitochondrial RNA and a complex of biologically active substances that provide anti-inflammatory and regenerative effects in damaged tissues.

To solve this problem, we propose a composition with anti-inflammatory and regenerative action, including

hyaluronic acid and / or its derivatives in an amount from 0.1 to 10 wt%,

fraction of mitochondrial RNA in an amount from 0.001 to 10 wt%,

saline or water - up to 100 wt%.

In the particular case of the invention, the composition additionally contains squalene in an amount of 0.01 to 10 wt%; taurine in an amount from 0.01 to 10 wt%; pantothenic acid (vitamin B5) in an amount from 0.01 to 10 wt%.

In the particular case of the invention, the composition is intended for injection, enteral, intra-articular and local administration.

The technical result of the claimed invention is as follows.

One of the types of biologically active molecules containing chains of nucleotides is mitochondrial RNA. The mitochondrial RNA fraction consists of linear biopolymers consisting of one nucleotide chain. RNA monomers (nucleotides) are composed of a five-carbon sugar - ribose, phosphoric acid residue and nitrogenous bases. Three nitrogenous bases in RNA molecules are the same as in DNA - adenine, guanine, cytosine, and the fourth is uracil. The mitochondrial fraction is 22 tRNA and 2 rRNA (16S and 23S), encoded by mitochondrial DNA. The fraction of mitochondrial RNA obtained from stem cells of avian or mammalian embryos does not differ in activity from each other, but differs from the fraction obtained, for example, from differentiated or germ cells. Fraction of mitochondrial RNA, being a modulator of DNMT activity, stimulates cell proliferation due to the modernization of the epigenome, oncoprotective (acts as an inhibitor of HDAC and HAT) and anti-inflammatory effect by stimulating the formation of the extracellular matrix (ECM) and activation of the healing ability of epithelial and connective tissue cells of the human body also restores the tissue's ability to regenerate, reduced with age.

In addition, the resulting composition of hyaluronic acid has not only good physical properties of viscoelasticity, but also resistance to hyaluronidase.

Additionally, the following biologically active components are added to the claimed composition: taurine (C ₂ H ₇ NO ₃ S), squalene (C ₃₀ H ₅₀₎ and pantothenic acid (C ₉ H ₁₇ NO ₅ ), which provide the following functions: taurine is a membrane stabilizer. It strengthens cell membranes and regulates their permeability to water and electrolytes. Thanks to this amino acid, such essential electrolytes as potassium and magnesium remain inside the cell, and sodium outside. This creates an osmotic gradient, the difference in electrolyte and water content on both sides of the cell membrane. Squalene is a powerful antioxidant and immunomodulator. Pantothenic acid is involved in the formation of coenzyme A, Coenzyme A ensures the most important biochemical reactions in our body. One of these reactions is the tricarboxylic acid (TCA) cycle. CTK provides cellular respiration, in which glucose is utilized under the action of oxygen to form carbon dioxide and energy in the form of ATP molecules. In the course of TCA, many of the most important organic compounds for us are utilized and synthesized: carbohydrates, fatty acids, amino acids. In turn, fatty acids and amino acids are the building blocks of fats (lipids) and proteins. Thus, coenzyme A, and, accordingly, pantothenic acid, regulates all the main links of carbohydrate, lipid and protein metabolism.

Disclosure of the essence of the invention.

The present invention relates to a composition containing hyaluronic acid and / or its derivatives, having a degree of crosslinking from 0.1 to 150%, and containing a fraction of mitochondrial RNA from 0.001 to 10% by weight based on the total weight of the composition.

The term "hyaluronic acid" as used in this application means a biopolymer material in which repeating units containing N-acetyl-D-glucosamine and D-glucuronic acid are connected in a linear fashion. In the present invention, the term hyaluronic acid is used to refer to the actual hyaluronic acid and its salt, for example the sodium salt - sodium hyaluronate.In the present invention, hyaluronic acid as such or its salt can be used alone or as a combination of the two. In the present invention, the molecular weight of hyaluronic acid may be 50,000 to 4,000,000 Da.

Derivatives of crosslinked hyaluronic acid can be obtained by crosslinking hyaluronic acid as such or its salt using a crosslinking agent. For crosslinking, you can use the method of using a crosslinking agent in an aqueous alkaline solution. The crosslinking agent can be an ether such as 1,4-butanediol diglycidyl ether (BDDE) or an aqueous alkaline solution, preferably NaOH. An aqueous solution of NaOH can be used at a concentration of 0.25 to 5N. Crosslinked hyaluronic acid derivatives are viscoelastic crosslinked hyaluronic acid. The term "degree of crosslinking" as used in this application means the% (wt.) Of the crosslinking agent relative to the monomer units of hyaluronic acid in the crosslinked part of the composition based on hyaluronic acid. The "degree of crosslinking" used in this application means the% (wt.) Of the crosslinking agent relative to the monomeric units of hyaluronic acid in the crosslinked part of the composition based on hyaluronic acid. The degree of crosslinking is defined as the weight ratio of the crosslinking agent to the weight ratio of the hyaluronic acid monomer. In the present invention, the degree of crosslinking of hyaluronic acid is preferably in the range of 1 to 50% when crosslinked with a crosslinking agent.

In addition, the composition of the present invention comprises mitochondrial RNA (mtRNA) fractions. In the present invention, the mitochondrial RNA fraction consists of linear biopolymers consisting of a single nucleotide chain.

The mitochondrial RNA fraction is obtained in the following way. Chicken embryos 4-8 days old were subjected to disaggregation with 0.05% trypsin solution by incubation in the solution for 10 minutes at 37 ° C. The resulting cell suspension was placed in test tubes and centrifuged at 100g for 10 minutes. The supernatant was removed, the pellet containing the cells was resuspended in hypotonic buffer (4 mM Tris-HCl pH 7.8, 2.5 mM NaCl, 0.5 mM MgCl2, and 2.5 mM PMSF) and incubated for 6 min on ice ... The swollen cells were destroyed in a homogenizer. Isotonic levels were restored by adding 1/10 of the resulting volume of hypertonic buffer (400 mM Tris-HCl pH 7.8, 250 mM NaCl and 50 mM MgCl2), centrifugation was performed at 1200 g, the pellet containing nuclei and cell fragments was removed Mitochondria were obtained by centrifugation of the remaining supernatant at 13000g. Mitochondria were further purified using a 1.0 M and 1.5 M sucrose gradient. After centrifugation for 20 min at 60,000 g, the mitochondrial layer was harvested, resuspended in PBS, exposed to UV light at 302 nm for 6 min in a ChemiDoc instrument (Biorad) and concentrated by centrifugation at 13000g. To isolate RNA, mitochondria are suspended at a temperature of 0-2 ° C in 0.01M potassium acetate buffer, pH = 5.1, containing 1% sodium dodecyl sulfate and a mixture of 0.5% polyvinyl sulfate and 0.5% macaloid. An equal volume of a phenol-chloroform mixture is added to the resulting suspension, shaken for 10 minutes, then centrifuged at 5000 g for 20 minutes at a temperature of 2 ° C. The lower layer is removed with a syringe, an equal volume of phenol-chloroform mixture is again added to the remaining volume and centrifuged at 5000 g for 20 minutes. The bottom layer is removed, the remaining layers are extracted with chloroform, shaking for 5 minutes and centrifuging at 5000 g for 15 minutes, removing the bottom layer. Mitochondrial RNA is then precipitated from the remaining upper layer by adding a double volume of cold ethanol containing 0.1 mol NaCl. The resulting mitochondrial RNA is purified by gel filtration chromatography and subsequent capillary electrophoresis.

Obtaining mitochondrial RNA is also possible in another known way known from the prior art.

Additionally, the composition of the present invention includes the addition of squalene in an amount of 0.01 to 10 wt%; taurine in an amount from 0.01 to 10 wt%; pantothenic acid (vitamin B5) in an amount from 0.01 to 10 wt%.

The composition production method includes the following stages:

1) obtaining crosslinked hyaluronic acid and / or its salt (as a derivative of hyaluronic acid) to a degree of crosslinking from 0.1 to 150% by dissolving it in water or 0.9% NaCl solution with the addition of an aqueous alkaline solution using crosslinking agents;

2) mixing a fraction of mitochondrial RNA, obtained, for example, according to the above method, into derivatives of hyaluronic acid having a degree of crosslinking from 0.1 to 150%,

In a further embodiment of the invention, the steps take place

3) mixing a biologically active complex consisting of squalene, taurine, pantothenic acid.

4) the step of packaging the product into the desired carrier, container or container (eg, a container for a pre-filled syringe).

Thus, the composition of a gene-activated hydrogel, consisting of hyaluronic acid, a fraction of mitochondrial RNA and, optionally, a biologically active complex: squalene, taurine, pantothenic acid according to the present invention can be used for cosmetic, prophylactic or therapeutic purposes due to its elasticity properties, resistance to the action of the enzyme and the ability to increase tissue regeneration. For example, such an injectable composition can be used as a composition for filling or replacing biological tissue, filling in wrinkles, remodeling the face, a composition for enhancing the lips, a composition for skin rehydration therapy with mesotherapy, a composition for replacing or temporarily filling synovial fluid. for arthritis, compositions for increasing the volume of the sphincter of the urethra, reducing the lumen of the vagina, in urology or gynecology, compositions for auxiliary action or treatment in the surgical treatment of cataracts in ophthalmology, pharmaceutical gel for the release of the active substance or compositions for restoring bones, increasing the volume of the vocal cords, education surgical tissue, injectable and external agent for the treatment of alopecia.

The composition is proposed to be administered in the form of intra-articular injections in a volume of 2-3 ml, subcutaneous injections at a dosage of 0.5-20 mg / ml, enterally at a dose of 2-5 mg / kg, or externally, for example, by topical application to the wound surface.

Brief description of explanatory materials.

Table 1. Wound healing in rats after thermal burn.

Table 2. Evaluation of the anti-inflammatory activity of the claimed composition.

Table 3. Comparative analysis of the viscosity of the claimed hydrogel and known.

Implementation of the invention

Example 1. Getting a composition.

Stage 1: Obtaining hyaluronic acid.

2 g of hyaluronic acid molecular weight: 2 - 2.5 million Da was mixed with 0.9% NaCl solution, 0.25N NaOH solution, BDDE was added in an amount of 0.5 ml to achieve a crosslinking degree of 25%, and crosslinked by keeping the mixture within 10 minutes at a constant temperature of 85 ° C. As a result, 95 ml of a base gel of a viscoelastic consistency was obtained.

Stage 2. Obtaining mitochondrial RNA. The mitochondrial RNA fraction is obtained from the mitochondria of chicken embryo cells in accordance with the method, for example, described above, purified by gel filtration chromatography and subsequent capillary electrophoresis of the supernatant of the embryonic cell culture of the chicken embryo. The result is a crystalline powder of mitochondrial RNA.

Stage 3. Preparation of a composition from hyaluronic acid and mitochondrial RNA.

Crystalline mitochondrial RNA powder in an amount of 0.2 mg is dissolved in 2 ml of 0.9% NaCl solution and ex tempore is added to the base cooled gel (see stage 1) with stirring under standard conditions at room temperature using a magnetic stirrer for 30 minutes.

Example 2. Obtaining a composition.

According to the invention, the claimed composition may additionally contain such biologically active substances as squalene, taurine, pantothenic acid. In this case, the preparation of the composition is carried out according to example 1, with the addition of a stage at which the addition of biologically active substances in low concentrations to the composition obtained in steps 1-3 is carried out: squalene 100 μg per 1 ml of 0.9% NaCl; taurine 400 μg per ml 0.9% NaCl; pantothenic acid 400 μg per 1 ml of 0.9% NaCl. All three bioactive substances are placed in a laboratory vacuum homogenizer HM-220VM manufactured by HT Machinery Japan. Squalene is homogenized to 1-5 micron drops and pantothenic acid and taurine are completely dissolved, then the resulting mixture is mixed into the composition using a DSMS-100 laboratory magnetic stirrer from Digisystem Taiwan.

Example 3. Hyaluronic acid with a molecular weight of 2 - 2.5 million Da was mixed with saline (0.9% NaCl), NaOH solution up to 10% (wt.) And BDDE was added to achieve a degree of crosslinking of 5%, and subjected to crosslinking at constant temperature 85 ° C. As a result, 99 ml of a base gel of a viscoelastic consistency was obtained. A composition from a gene-activated gel containing 0.5 mg / ml was obtained by adding, to the base gel, a solution containing a mitochondrial RNA fraction in an amount of 5 mg per 1 ml of saline (0.9% NaCl). This composition has been used as a gel for filling facial wrinkles and improving skin tissue regeneration.

Example 4. The sodium salt of hyaluronic acid with a molecular weight of 2 - 2.5 million Da was mixed with saline (0.9% NaCl), NaOH solution up to 10% (wt.) And ether was added to achieve a degree of crosslinking of 5%, and subjected to crosslinking at a constant temperature of 85 ° C. As a result, 99 ml of a base gel of a viscoelastic consistency was obtained. A composition from a gene-activated gel containing 0.5 mg / ml of mitochondrial RNA fraction was obtained by adding to the base gel a solution containing a mitochondrial RNA fraction in an amount of 5 mg per 1 ml of physiological solution (0.9% NaCl). gel for filling facial wrinkles and regenerating skin tissues.

Example 5. Hyaluronic acid with a molecular weight of 2-2.5 million Da was mixed with saline (0.9% NaCl), NaOH solution up to 10% (wt.) And ether was added to achieve a degree of crosslinking of 2.5%, and subjected to crosslinking at a constant temperature of 85 ° C. As a result, 98 ml of a base gel of a viscoelastic consistency was obtained. A composition from a gene-activated gel containing 0.5 mg / ml was obtained by adding to the base gel a solution containing a mitochondrial RNA fraction in an amount of 5 mg per 1 ml of physiological solution (0.9% NaCl), then a complex of biologically active substances with a total weight of 5 mg consisting of the squalene isomer; L-Taurine; in a ratio allowing to achieve complete dissolution in 1 ml of solvent and addition of 1 mg of Pantothenic acid. The specified composition was used as a gel for biorevitalization and regeneration of facial skin tissues.

Example 6. Hyaluronic acid with molecular weight: 2.5-3 million Da was mixed with saline (0.9% NaCl), NaOH solution up to 10% (wt.) And ether was added to achieve a degree of crosslinking of 5%, and subjected to crosslinking at constant temperature 85 ° С. As a result, 99 ml of a base gel of a viscoelastic consistency was obtained. A composition from a gene-activated gel containing 0.5 mg / ml was obtained by adding to the base gel a solution containing a mitochondrial RNA fraction in an amount of 5 mg per 1 ml of physiological solution (0.9% NaCl), after which the microspheres of cross-linked hyaluronic acid were mixed until degree of stitching 1000% and sizes from 1 to 999 nm. The specified composition was used by injection as an implant replacing the synovial fluid of the knee joint. The introduction of microspheres made it possible to drastically reduce the friction of the articular surfaces by converting sliding friction into rolling friction. And the regeneration of the cartilaginous connective tissue of the joint.

Example 7. Hyaluronic acid with a molecular weight of 3-3.3 million Da was mixed with saline (0.9% NaCl), As a result, 99 ml of a base gel of a visco-elastic consistency was obtained. A composition from a gene-activated gel containing 0.5 mg / ml was obtained by adding to the base gel a solution containing a mitochondrial RNA fraction in an amount of 5 mg per 1 ml of physiological solution (0.9% NaCl) The specified composition was used injections as an implant gel for the treatment of periodontal disease, namely the closure of the gum of the exposed neck of the tooth. regeneration of periodontal connective tissue.

Example 8. Hyaluronic acid with a molecular weight of 3-3.3 million Da was dissolved in water, as a result, 99 ml of a base gel of a visco-elastic consistency was obtained. A composition from a gene-activated gel containing 0.5 mg / ml was obtained by adding a solution containing a mitochondrial RNA fraction in an amount of 5 mg per 1 ml of saline (0.9% NaCl) to the base gel. active substances with a total weight of 5 mg consisting of the squalene isomer; L-Taurine; in a ratio that allows to achieve complete dissolution in 1 ml of solvent and 1 mg of pantothenic acid, this composition was used as a biorevitalizant for the treatment of diffuse alopecia, regeneration of skin tissues and hair follicles.

Example 9. Hyaluronic acid with a molecular weight of 0.5-4 million Da was mixed with saline (0.9% NaCl), As a result, 85 ml of a base gel of a visco-elastic consistency was obtained. A composition from a gene-activated gel containing 2.5 mg / ml mitochondrial RNA fraction was obtained by adding to the base gel a solution containing a mitochondrial RNA fraction in an amount of 50 mg per 5 ml of physiological solution (0.9% NaCl), then a complex of biologically active substances with a total mass of 50 mg consisting of the squalene isomer; L-Taurine; in the ratio allowing to achieve complete dissolution in 10 ml of solvent and additives of 10 mg of Pantothenic acid, the specified composition was used for the treatment of gastritis to relieve inflammation, regeneration of the gastric epithelium.

Example 10. Hyaluronic acid with a molecular weight: 0.5-4 million Da was mixed with saline (0.9% NaCl), as a result, 85 ml of a base gel of a visco-elastic consistency was obtained. A composition from a gene-activated gel containing 2.5 mg / ml mitochondrial RNA fraction was obtained by adding to the base gel a solution containing a mitochondrial RNA fraction in an amount of 50 mg per 5 ml of saline (0.9% NaCl) .The composition was used to treat cystitis for removal of inflammation of the regeneration of the epithelium of the bladder using a urethral catheter.

Example 11. Confirmation of the reparative properties of the claimed composition.

The reparative activity of the claimed composition in various versions of its execution was carried out on a model of a burn wound.

The studies were carried out on white outbred male rats with a total body weight of 310 ± 10 g. A series of experiments were carried out on 50 animals from 5 groups: control and 4 experimental groups:

Group 1 - control,

2 - a composition containing 0.1 wt% hyaluronic acid and 0.001 wt% mitochondrial RNA, 0.9% NaCl solution up to 100 wt% was applied to the burn surface,

3 - a composition containing 10 wt% hyaluronic acid, 10 wt% mitochondrial RNA, water - up to 100 wt% was applied to the burn surface,

4 - a composition containing 0.1 wt% sodium hyaluronate, 0.001 wt% mitochondrial RNA and squalene in an amount of 0.01 wt% was applied to the burn surface; taurine in an amount of 0.01 wt%; pantothenic acid (vitamin B5) in an amount of 0.01 wt%,

5 - applied to the surface of the burn a composition containing 10 wt% hyaluronic acid, 10 wt% mitochondrial RNA and squalene in an amount of 10 wt%; taurine in an amount of 10 wt%; pantothenic acid (vitamin B5) in an amount of 10 wt%.

The control group of animals did not apply any preparations to the burn surface; the healing of burn wounds took place independently.

A burn wound was simulated using an electric device for a contact thermal burn, the area of the contact metal plate was 214 mm ² , the application time was 10 seconds, the degree of burn was IIIa, the average for all groups, the initial area of each on the 1st day 232.8 ± 6.5 mm ² , total - about 15.0% of the body surface area. Each animal received 2 symmetrical burns on the previously depilated areas of the skin of the posterolateral surface of the body on both sides of the spine. In the experimental groups, one site was a control: The second site was an experimental one, on its surface every day, once a day during the entire observation period, the studied variants of the composition were applied in accordance with the groups. The burn area was measured in a planimetric manner. Measurements were made on the 1st day after the application of the burn and then every 5 days until complete healing. The results are presented in table. 1.

Example 12. Confirmation of the anti-inflammatory effect of the claimed composition.

The anti-inflammatory effect of the claimed composition was studied in the model of carrageenan edema of the paw in rats. Acute inflammatory reaction is reproduced by subplantar injection of 0.1 ml of 1% carrageenan solution. The severity of the inflammatory response after the induction of inflammation is assessed by the change in the paw volume (oncometrically).

The experiment was carried out on male Wistar rats weighing 160-220 g in the amount of 90 individuals. The animals were divided into 9 groups of 10 individuals - control and 8 experimental:

1 - control, a saline solution was administered orally,

2 - received orally a composition containing 0.1% wt% hyaluronic acid and 0.001 wt% mitochondrial RNA,

3 - received orally a composition containing 10 wt% hyaluronic acid, 10 wt% mitochondrial RNA,

4 - received orally a composition containing 0.1% wt% hyaluronic acid, 0.001 wt% mitochondrial RNA and squalene in an amount of 0.01 wt%; taurine in an amount of 0.01 wt%; pantothenic acid (vitamin B5) in an amount of 0.01 wt%,

5 - received orally a composition containing 10 wt% hyaluronic acid, 10 wt% mitochondrial RNA and squalene in an amount of 10 wt%; taurine in an amount of 10 wt%; pantothenic acid (vitamin B5) in an amount of 10 wt%,

6 - received a topically applied composition containing 0.1% wt% hyaluronic acid and 0.001 wt% mitochondrial RNA,

7 - received a topically applied composition containing 10 wt% hyaluronic acid, 10 wt% mitochondrial RNA,

8 - received topically applied composition containing 0.1% wt% hyaluronic acid, 0.001 wt% mitochondrial RNA and squalene in an amount of 0.01 wt%; taurine in an amount of 0.01 wt%; pantothenic acid (vitamin B5) in an amount of 0.01 wt%,

9 - received a topically applied composition containing 10 wt% hyaluronic acid, 10 wt% mitochondrial RNA and squalene in an amount of 10 wt%; taurine in an amount of 10 wt%; pantothenic acid (vitamin B5) in an amount of 10 wt%.

In the experimental groups, the composition was administered 1 hour before the induction of inflammation, and the volume of the limb was measured before the induction of inflammation and 3 hours and 24 hours after the induction. The composition was orally administered through a tube intragastrically in a volume of 2 ml, the control group was injected with 2 ml of saline. Topically, the composition was applied to the surface of the inflamed limb with an applicator stick.

As follows from table 2, the use of the claimed composition reduces the severity of inflammation when used topically and orally.

Example 13. Study of the stability of the claimed composition to the effects of hyaluronidase.

A comparative analysis of the resistance of the claimed composition in various variants of its embodiment and the known product for tissue augmentation Restylane® to the effect of hyaluronidase was carried out. A hyaluronidase solution (15 μl of a 0.15 mg / ml hyaluronidase solution in 1.9 mM phosphate buffer, ≈2.5 units) is added to 0.75 g of each of the embodiments of the present invention and mixed thoroughly. Each mixture is loaded into a syringe and centrifuged for 1 min at 1000 rpm to remove air bubbles. Then 0.35 g of each mixture is placed on a rheometer plate. The viscosity of the gel is recorded every 30 minutes for 5 hours at 37 ° C. on a flat plate, with a gap of 200 μm and a frequency of 1 Hz. Determine the dependence of the elastic modulus G '(Pa) on time. The results are presented in table. 3

As can be seen from the data given in table. 3, the claimed composition in various versions of its execution has a high resistance to hyaluronidase, exceeding that of other known and used in practice hydrogels.

Thus, the claimed composition has an anti-inflammatory and regenerative effect, as well as a pronounced resistance to the effects of hyaluronidase and can be used in medical and cosmetic practice.

To the application " Composition containing the epigenome modernization factor, which has a reparative, oncoprotective and anti-inflammatory effect "

Burn area on the 5th day, mm ² Burn area on the 10th day, mm ² 1st group 178.3 ± 2.2 155.6 ± 1.9 Group 2 154.2 ± 1.9 125.4 ± 2.1 Group 3 164.1 ± 1.3 136.7 ± 1.8 4 group 168.7 ± 0.9 135.9 ± 2.3 Group 5 152.3 ± 1.6 126.2 ± 1.5

Table 1.

To the application " Composition containing the epigenome modernization factor, which has a reparative, oncoprotective and anti-inflammatory effect "

The increase in the volume of the paws,% in relation to the original after the introduction of carrageenan After 3 h After 24 h 1st group 40 ± 2.7 38 ± 1.8 Group 2 16.5 ± 1.6 23.4 ± 2.4 Group 3 20.1 ± 1.8 25.3 ± 1.8 4 group 18.3 ± 2.3 23.6 ± 2.1 Group 5 19.9 ± 1.9 22.9 ± 3.1 6 group 17.5 ± 2.2 23.2 ± 2.2 7 group 21.5 ± 1.6 24.8 ± 1.9 8 group 17.4 ± 1.5 23.1 ± 2.1 Group 9 20.7 ± 2.6 25.5 ± 2.3

Table 2.

To the application " Composition containing the epigenome modernization factor, which has a reparative, oncoprotective and anti-inflammatory effect "

sample Incubation time, min 30 minutes 60 minutes 180 minutes 300 minutes Restylane® 320 300 200 100 The claimed hydrogel with 0.1 wt% hyaluronic acid and 0.001 wt% mtRNA 400 360 320 300 The claimed hydrogel with 10 wt% hyaluronic acid and 10 wt% mtRNA 450 435 418 405 The claimed hydrogel with 0.1 wt% hyaluronic acid and 0.001 wt% mtRNA + squalene 0.01 wt%; taurine 0.01 wt%; vitamin B5 at 0.01 wt%. 410 390 375 362 The claimed hydrogel with 10 wt% hyaluronic acid and 10 wt% mtRNA + squalene 10 wt%; taurine 10 wt%; vitamin B5 10 wt%. 445 430 417 403

Table 3.

Claims (5)

1. Composition with anti-inflammatory and regenerative action, including hyaluronic acid and / or its derivatives in an amount of 0.1 to 10 wt%, a fraction of mitochondrial RNA in an amount of 0.001 to 10 wt%, 0.9% NaCl solution or water up to 100 wt%. 2. The composition according to claim 1, characterized in that it additionally contains squalene in an amount from 0.01 to 10 wt.%; taurine in an amount from 0.01 to 10 wt%; pantothenic acid (vitamin B5) in an amount of 0.01 to 10 wt%. 3. Composition according to PP. 1, 2, intended for injection, intra-articular, enteral and local use. 4. Composition according to PP. 1, 2, intended for intra-articular use, containing microspheres of crosslinked hyaluronic acid up to the degree of crosslinking 1000%, where the degree of crosslinking is% (wt.) Of the crosslinking agent relative to the monomer units of hyaluronic acid in the crosslinked part of the composition based on hyaluronic acid and sizes from 1 up to 999 nm.