RU2730477C1 - Method for producing biologically active pharmaceutical substance for feeding extracellular matrix and preventing locomotor diseases and barrier functions of respiratory and digestive organs - Google Patents

Abstract

FIELD: medicine; pharmaceuticals.SUBSTANCE: invention refers to pharmaceutics, namely to a method for preparing a biologically active pharmaceutical substance for feeding an extracellular matrix and preventing locomotor diseases and barrier functions of respiratory and digestive apparatus. Method involves preparation of raw material - mechanical activation and grinding to a certain extent in a medium of inert gas or carbon dioxide, vegetable silicon-containing material, chondroitin sulphate, glucosamine sulphate, hyaluronic acid, methylsulphonylmethane, mixing components in certain proportions to homogeneous state, repeated mechanical activation of the obtained mixture in the medium of inert gas or carbon dioxide, milling to particle size of 1–5 mcm and microcapsules in cellulose microstructures.EFFECT: method provides creating a biologically active pharmaceutical substance for feeding an extracellular matrix and preventing locomotor diseases and barrier functions of respiratory and digestive organs.4 cl

Description

A method of obtaining a biologically active, pharmaceutical substance for nutrition of the extracellular matrix and prevention of diseases of the musculoskeletal system and barrier functions of the respiratory and digestive organs.

IPC A61K 36/899, A61K 31/10, A61K 31/415, A61K 31/726, A61P 19/02

The invention relates to the food, pharmaceutical industry and experimental medicine, in particular, to the production of drugs for therapeutic and prophylactic purposes for the musculoskeletal system, the mucous membrane of the respiratory and digestive organs of humans in the form of a biologically active substance for the preparation of transdermal systems.

The object of influence and restoration is the extracellular matrix (ECM) of human body tissues, and the technical solution is aimed at providing nutrition and restoration of the extracellular matrix responsible for the functionality of the cartilaginous and connective tissue of the musculoskeletal system, mucous membranes of the respiratory and digestive organs, which provide barrier functions from external bacterial and viral effects.

The extracellular matrix is a multidimensional structure that surrounds and supports tissue cells. This structure has a diffuse mode of nutrition and regeneration and provides mechanical support for cells and transport of chemicals. ECM can be conditionally referred to as connective tissue. It consists of three main types of molecular structures such as protoglycans, fibrillins, and fibronectins. For general purposes, we classify them as biopolymers.

In recent publications, the role of EMC is considered in a number of antitumor processes in the body http://www.dslib.net/citologia/funkcionalnaja-rol-sistemy-biosinteza-geparansulfata-v-kancerogeneze.html

To nourish the matrix, the body uses a number of molecules such as chondroetins and glucosamines, hyluaronic acid, mitylsulfonylmethane (MSM) and biophilic silicon. The last two are responsible for the rate of chemical reactions in tissue regeneration.

Collagen is a protein that is the main component of the extracellular matrix and an integral part of connective tissue. Collagen, when implanted in tissue, stimulates reparative processes and is a biodegradable material.

Known patent RU No. 2249462, published 10.04.2005. which offers a biopolymer matrix for the regeneration of body tissues.

The basis of this biopolymer matrix is a heterogeneous collagen-containing structure consisting of phases: solid - in the form of microparticles (microspheres) from the collagen of mammalian tissue, and liquid - from denatured collagen of avian tissue with the phase ratio (1-10) :( 1-10). The matrix (at room temperature) is an elastic mass (gel) and has a microparticle size of 100-300 microns. The heterogeneous structure of the matrix allows to increase and regulate the time of collagen biodegradation, which increases the efficiency of tissue repair. Additionally, the matrix may contain additives that promote the growth of cells and tissues, in particular, cells of nerve tissues. According to the invention, the advantages of this matrix are: biocompatibility; support-trophic function and the ability to stimulate cell proliferation.

On the basis of this technical solution, a biopolymer matrix was created for the proliferation and regeneration of tissues based on a heterogeneous collagen-containing structure consisting of collagen microparticles with a size of 30-300 microns, distributed in a homogeneous collagen gel at a phase ratio (1-10) :( 1-10), with swelling capacity not less than 87%, pH 4.8-7.2 (commercial product "Sphero®GEL", CJSC "Biomir-service", Russia).

At the same time, studies of this heterogeneous collagen-containing structure when using it as an implantable neuroendoprosthetic system in a defect in nerve tissue have shown that its regenerating potential in relation to nerve tissues is not fully realized.

Known biopolymer matrix for cell proliferation and tissue regeneration, which is developed on the basis of a heterogeneous collagen-containing structure consisting of collagen microparticles with a size of 30-300 microns, distributed in a homogeneous collagen gel, and containing neural stem cells and a stimulating additive in the form of silicon nanoparticles with a size of 50 -150 nm in an amount of 1⋅10-3-1⋅10-2 wt. %. The nanoparticle silicon additive is used to stimulate cell proliferation and nerve tissue regeneration. Silicon nanoparticles are obtained by laser-induced pyrolysis of monosilane SiH4. In vitro evaluation of the biological effect of the biopolymer matrix (influence on cell viability and proliferation) was carried out using neural stem cells. The biopolymer matrix simulates cell proliferation and regeneration of nerve tissues and has biocompatible properties (RU 2478398, IPC A61K 38/39, A61K 35/12, В82В 1/00, A61K 47/02, А61Р 25/00, 04/10/2013). When obtaining a building substance for the recovery of the extracellular matrix, the regeneration of VM in combination with silicon nanoparticles, a derivative of nitrogen obtained from selan is used as a stimulator of absorption.

Silicon particles are used in cosmetology, medicine, orthopedics and biocompatible access (see - RU No. 2375080, publications. 27.03.2009; RU No. 2003137823, publications. 27.03.2005; USA No. 7186287, publications. 06.03.2007; WO. 2010 / 096733 A2, publ., 26.08.2010; L. A. Osminkina, E. N. Lukyanova, et al. Effect of nanostructured nitrogen on the proliferation of stem and cancer cells - Bulletin of Experimental Biology and Medicine. Russian Academy of Medical Sciences. Moscow, 2011, Volume 151, No. 1, pp. 91-95).

Nanoparticles of nitrogen are obtained on the basis of the technologies of laser-induced pyrolysis of monosilane SiH ₄ , described in the works: A.G. Vladimirov, S.B. Korovin, V.I. Empty. Luminescence of silicon nanoparticles. Institute of General Physics named after A.M. Prokhorov RAS. Collection of abstracts of the Third International Forum on Nanotechnology RUSNANOTECH. Moscow, 2008, p. 767-770; A. Vladimirov, S. Korovin, A. Surkov, E. Kelm, V. Pustovoy. Institute of General Physics. AM Prokhorov RAS. "Synthesis of luminescent Si nanoparticles using laser-induced pyrolysis", Laser Physics, 2011, volume 21, no. 4, p. 830-835.

The process of obtaining nitrogen nanoparticles consists in feeding a reactive gas mixture into a flow reactor: monosilane (SiH ₄ ) and an added gas - helium (He), when the pyrolysis reactions of the gas mixture are indicated in continuous CO ₂ radiation - a laser when exposed to the gas mixture in the reactor is below atmospheric. Argon (Ar) uses a buffer gas for exothermic pyrolysis reactions. Under the influence of CO ₂ - laser radiation (output power 70 W, wavelength λ = 10.6 μm), SiH ₄ molecules decompose into active SiH _x precipitates, followed by silicon-silicon (Si-Si) interaction, which leads to the synthesis of nanoparticles in reaction zone.

Typical controlled parameters of the process (reaction): gas and monosilane: helium: argon, as 1: 1: (45-65); digital power of laser radiation 5000-8000 W / cm ² ; the pressure of the gas mixture in the reactor is 200-650 torr. In the range from 5 to 200 nm. The controlled process allows you to control the average size of nitrogen nanoparticles and obtain (if necessary) nanoparticles with a narrow size dispersion.

Obtained nitrogen nanoparticles. For research, a transmission (transmission) electron microscope TEM "LEO912 AB OMEGA" is used.

It should be noted that laser-induced pyrolysis of monosilane SiH ₄ makes it possible to obtain "clean surfaces" of nanoparticles, without any chemicals, reaction products and acid residues, depending on the parameters of electrochemical etching of the nitrogen plate (see the above-mentioned patent RU No. 2375080). In addition, according to research, nanoparticles have the property of biodegradation (for example, see - Ji-Ho Park, Luo Gul et al. Biodegradable luminescent porous silicon nanoparticles for in vivo applications. Nature Materials, 2009, 8, pp. 331-336).

Known composition with anesthetic,

anti-inflammatory and improving the functional state of the musculoskeletal system, containing glucosamine sulfate, chondroitin sulfate, methylsulfonylmethane, L-proline, ascorbic acid (vitamin C), magnesium, characterized in that it additionally contains zinc gluconate, citric acid, sweetener, flavor, preservative, water with the following ratios, wt. %: Glucosamine sulfate 1-20, Chondroitin sulfate 1-20, Methylsulfonylmethane 0.0001-10, L-proline 0.01-5, Ascorbic acid (Vitamin C) 0.1-10, Zinc gluconate 0.01-3,

Magnesium gluconate 0.05-2, Citric acid 0.1-2, Sweetener 0.001-1.5, Flavoring 0.001-0.5, Preservative 0.01-1.0, Water Remaining up to 100 (RU 2468805, IPC A61K 31 / 737, A61K 31/7008, A61K 31/10, A61K 31/401, A61K 31/375, A61K 31/191, A61K 33/30, A61K 33/06, A61P 19/00, 10.12.2012).

The disadvantage of this patent is that bioavailability is provided only due to the liquid form used orally. The possibility of creating transdermal preparations (ointments and long-term patches) is complicated.

A known composition with analgesic and anti-inflammatory action, containing chondroitin sulfate, phenyltrimethicone, is stable 30, cetyl alcohol, cetyl palmitate, carbopol, solubilizer, triethanolamine, glucosamine hydrochloride, liposomes, beta-cyclodextolands, essential oil of nanodextrin, essential oil no hyaluronidase, ginger extract, harpagophytum extract, euxil K 727, demineralized water, at a certain ratio of components. A topical composition with analgesic and anti-inflammatory action, containing chondroitin sulfate, phenyltrimethicone, stable 30, cetyl alcohol, cetyl palmitate, carbopol, solubilizer, triethanolamine, glucosamine hydrochloride, liposomes, beta-cyclodextrin, essential oil, ericin, essential oil betulin, hyaluronidase, ginger extract, harpagophytum extract, euxil k 727, demineralized water, at a certain ratio of components. The above compositions have a pronounced analgesic and anti-inflammatory effect, are stable (RU 2539390, IPC A61K 36/9066, A61K 31/724, A61K 31/737, 01.20.2015).

The disadvantage of this patent is that the technical solution has only the possibility of dermal application, it contains many substances that irritate the skin to increase bioavailability.

Known combined active composition for the treatment of diseases of the musculoskeletal system, which is a pharmaceutical composition of active substances for external use, made in the form of a soft dosage form. A bifunctional drug is described, containing chondroitin sulfate, celecoxib, dimethyl sulfoxide and an ointment base as biologically active substances - the rest, at a certain ratio of components. The tool provides the creation of a drug for transdermal use in the form of a gel for long-term local use, which has reparative, anti-inflammatory and analgesic effects and can be used as an independent or auxiliary drug in conditions associated with a decrease in the functions of the musculoskeletal system (RU 2582974, IPC A61K 31/10, A61K 31/415, A61K 31/726, A61P 19/02, 04/27/2016).

The disadvantage of this patent is that it is a transdermal product, its substance cannot be used orally. The assimilation is based on the properties of the protic solvent Dimexide, which is toxic. The onset of action on the body, as well as the two mentioned above, is about two months from the beginning of the intake.

In addition, we consider it necessary to indicate the most common basic disadvantages of competitors:

- Glucosamine and its salts are unstable in aqueous solutions and undergo rapid degradation. The technology for obtaining this product uses the dissolution of glucosamine in distilled water when heated, which accelerates the destruction process. This fact is confirmed by the fact that on the pharmaceutical market there are drugs and dietary supplements containing glucosamine only in solid form (tablets, capsules, lyophilisate).

- Non-selective NSAIDs are used as substances that relieve inflammation and pain, the main disadvantage of which is the inhibition of the synthesis of both forms of cyclooxygenase: COX-1, which is constantly synthesized in the body and is responsible for the normal course of physiological processes, and COX-2, synthesized only with the development of pathogenic factors leading to the development of inflammatory reactions. This fact explains the occurrence of serious side effects when using non-selective NSAIDs (gastropathology, anemia, instability of blood pressure).

We consider it necessary to rely on biophilic (chelated) silicon as a catalyst for assimilation. Its role in calcium absorption is noted in many sources, for example, RU 2397663 (drink).

The disadvantage of this known drink is the presence in it in a sufficiently large amount of calcium in the complete absence of silicon, although it is known that it is silicon that regulates the absorption and proper distribution of calcium in the body.

Based on the above, it is obvious that the problem lies in the need to ensure different forms of use of a biologically active, pharmaceutical substance, both for adjusting the human diet, and for a prophylactic or therapeutic drug with dosages ranging from 1 to 5 mg of the finished product per 50 kg of human weight. ...

The objective of the invention is to create a product for nutrition of the human extracellular matrix in the form of a biologically active pharmaceutical substance consisting of nutritional ingredients in a bioavailable form in a composition with assimilation catalysts - biophilic silicon and MSM (mitylsulfonylmethane) for use as a food additive for humans and a substance for preparation transdermal systems.

The technical result consists in creating a biologically active, pharmaceutical substance for adjusting the human diet or other forms of use as a preventive drug for diseases of the musculoskeletal system and barrier functions of the respiratory and digestive organs.

The technical result is achieved by a method of obtaining a biologically active, pharmaceutical substance for nutrition of the extracellular matrix and the prevention of diseases of the musculoskeletal system and barrier functions of the respiratory and digestive organs, including the preparation of raw materials, the introduction of additives and microencapsulation into the microstructures of cellulose, characterized by the fact that they are preliminarily subjected to mechanoactivation and grinding to size

10-100 microns in an inert gas or carbon dioxide environment of plant siliceous raw materials, as well as separately - to the size of microparticles 50-100 microns chondroetin sulfate, glucosamine sulfate, hyaluronic acid, mitylsulfonylmethane (MSM). Further, additives in the proportion of 1 part of chondroetin sulfate 1 part of glucosamine sulfate 0.3 part of hiluaronic acid and 0.3 part of MSM are mixed until homogeneous with the mentioned previously prepared plant siliceous raw material in an amount of 50% by weight of the mixture, after which the resulting mixture is mechanically activated again in medium of inert gas or carbon dioxide at a temperature of 40C ° and crushed to a particle size of 1-5 microns.

In addition, mechanical activation is carried out at acceleration of grinding bodies of 100-250 m / s ² and a residence time in the processing zone of 5-20 seconds.

In addition, it is preferable to use a vegetable siliceous raw material in the form of embryonic shells of rice and / or barley and / or wheat.

In addition, repeated grinding of the substance to a particle size of 1-5 microns is carried out in an amount of 70% of the total mass.

The product obtained by the method is developed on the basis of a mixture of microparticles of the indicated elements obtained as a result of mechanical activation of the feedstock in flow-type mechanical activator mills at a temperature of 40 g. C. and the time spent in the zone of shear deformation from 5 to 20 seconds in an inert gas or carbon dioxide. Honroetin sulfate, glucosamine sulfate, hyaluronic acid, MSM are mechanically activated separately. After the preparation of microparticles with a dimension of 50-100 microns, they are mixed into mixers and a ready-made mixture of additives consisting of proportions - 1 part of chondroetin sulfate 1 part of glucosamine sulfate 0.3 part of hiluaronic acid and 0.3 part of MSM are mixed in an amount of 50% of the total mass of the mixture until homogeneous states with plant siliceous raw materials -

preferably embryonic shells of rice, barley and wheat, previously subjected to mechanical activation and grinding to a size of 10-100 microns. Then the mixture of ingredients is mechanically activated again in an inert gas or carbon dioxide environment at temperatures up to 40 g. C and is crushed to a particle size of 1-5 microns of about 70% of the total mass. In all cases, mechanical activation is carried out with the acceleration of the grinding bodies, which ensures the acceleration of the grinding bodies of 100-250 m / s ² and the residence time in the processing zone is 5-20 seconds.

In contrast to the known methods of obtaining a building substance for the recovery of the extracellular matrix in combination with silicon as a stimulator of assimilation, for example, patent RU 2478398, where a derivative of nitrogen obtained from selane is used as a stimulator and the regeneration of EMC is stimulated by silicon nanoparticles, its bundles are formed in the claimed product ... The size of these bundles is from 1 to 5 microns, while silicon and product ingredients, microencapsulated in cellulose, correspond to the nanosize of 50-150 nm.

The key difference is the object of influence: not the conditional cartilage and bones mentioned in analogs, but the extracellular matrix, which is a conductor to the cartilaginous and ligamentous tissues. They receive nutrients from EMC through diffusion. Therefore, the bioavailability of the assembly is a critical factor. The claimed method increases bioavailability due to the selected mode of mechanochemical action in a mechanoactivator in an inert gas environment. The second decisive factor is the availability of a catalyst for assimilation - chelated silicon. The entire assembly is packed into a delivery vehicle - cellulose, which is dimensioned to provide close contact in the areas of intestinal absorption.

The analogs mentioned in the prior art, in particular patent RU 2468805, provide bioavailability due to the liquid form, and the drug is administered orally. In the claimed method, bioavailability is increased due to preliminary joint mechanical destruction, when mutual adhesion occurs in the molecules of substances and deformations accumulate, which can significantly improve assimilation. In addition to the oral form, transdermal preparations (ointments and long-term patches) can be created on the basis of the substance obtained by the claimed method.

The product of the known patent RU 2539390 has only the possibility of dermal application and contains many skin irritating substances to increase the availability. In the product according to the claimed method, this is achieved due to the dimension of the active compounds that penetrate into the distance between the cells of the epidermis. In patent RU 2582974, a transdermal product is obtained, its substance cannot be used orally. The assimilation is based on the properties of the protic solvent Dimexide and is therefore toxic. For our product, the absorption catalyst is chelated silicon. It is a natural catalyst for building extracellular matrix, bone tissue and cartilage.

As mentioned above, the onset of the action of products of known analogues on the body is about two months from the beginning of the intake. The effect of the product of the proposed method is observed after two weeks.

Thus, the claimed method achieves a technical result, which consists in creating a biologically active, pharmaceutical substance for a food additive for adjusting the human diet or other forms of use as a prophylactic or therapeutic drug for diseases of the musculoskeletal system and human barrier functions.

Claims (4)

1. A method of obtaining a biologically active pharmaceutical substance for nutrition of the extracellular matrix and prevention of diseases of the musculoskeletal system and barrier functions of the respiratory and digestive organs, including the preparation of raw materials, the introduction of additives and microencapsulation into the microstructure of cellulose, characterized by the fact that it is preliminarily subjected to mechanical activation and grinding to size 10-100 microns in an inert gas or carbon dioxide environment of plant siliceous raw materials, as well as - separately - to the size of microparticles of 50-100 microns chondroetin sulfate, glucosamine sulfate, hyaluronic acid, mitylsulfonylmethane (MSM), then additives in a proportion of 1 part chondroetin sulfate, 1 part of glucosamine sulfate, 0.3 part of hyluaronic acid and 0.3 part of MSM are mixed until homogeneous with the above-mentioned prepared plant siliceous raw material in an amount of 50% of the mass of the mixture, after which the resulting mixture is mechanically reactivated in an inert gas or carbon dioxide medium at temperature of 40 ° C and crushed to a particle size of 1-5 microns. 2. The method according to claim 1, characterized in that the mechanical activation is carried out with the acceleration of the grinding bodies of 100-250 m / s ² and the residence time in the processing zone of 5-20 seconds. 3. A method according to claim 1, characterized in that the plant uses siliceous raw material in the form of embryonic shells of rice and / or barley and / or wheat. 4. The method according to claim 1, characterized in that the repeated grinding of the substance to a particle size of 1-5 microns is carried out in an amount of 70% of the total mass.