RU2763171C1 - Method for producing an emulsion containing nano- and microencapsulated biologically active substances - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention is aimed at creating a composition wherein an agent based on a strain of the Pleurotus ostreatus fungus RNCIM F-819 with polyfunctional biomedical activity is used as a biologically active substance, impacting the tissue metabolism. For prolonged effect, the agent is encapsulated in a shell constituting spherical colloidal particles of calcium carbonate CaCO₃. The particles are produced when mixing solutions of calcium chloride CaCl₂ and soda (or sodium carbonate) Na₂CO₃ with a concentration of 0.33 M.

EFFECT: composition provides enhanced therapeutic and preventive effect on human skin cover in the affected areas and preservation of this effect for a long time.

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Description

The invention relates to the creation of a composition intended for direct contact with the skin and use in medicine or at home, namely, the creation of therapeutic and prophylactic agents with natural ingredients for external use with a bactericidal, anti-inflammatory, anti-decubitus and anti-burn effect.

Currently, due to complex diseases, including surgical ones, there is a large number of bedridden patients with different etiologies. In medicine, the demand for drugs for the treatment of bedsores and burns is growing rapidly.

Bedsores occur for three reasons:

- from constant pressure,

- from heat and humidity,

- from perspiration.

Modern scientific research is directed, unfortunately, mainly to the development of new methods of treatment of already formed decubitus ulcers. Treatment of bedsores, and more importantly, prevention of bedsores, should be started as quickly as possible. Bedsores are associated with significant pain.

Recently, there has also been an increasing amount of inflammatory and burn effects on the skin. Therefore, the prevention and treatment of inflammatory and burn diseases of the skin requires a scientific and convenient everyday approach to solving this issue.

There are data confirmed in the literature that the use of traditional antiseptics (hexachlorophene, chlorhexidine, povidone-iodine, etc.) in the area of altered skin. destroys the cell membrane and thus slows down regeneration processes [Edlich, R.F., Rodeheaver, G.T., Kuphall, J., deHoll, J.D., Smith, S., Bacchetta, C.A., Edgerton, M.T. Technique of closures: Contaminated wound. //J. Am. Coll. Emerg. Phys. 3:375, 1974]. Therefore, more and more often, natural biologically active substances (BAS) are being used to treat skin lesions.

For the treatment of bedsores, vegetable fatty oils containing carotenoids (sea buckthorn, mountain ash, rosehip, and others), which have a wound healing ability, are often used as biologically active substances. [Collection of folk medicine and non-traditional methods of treatment, G.Z. Minejyan, M., 1991].

An anti-decubitus oil is known containing fatty oil of sea buckthorn, wheat germ, tamanu oil and essential oils of oregano, geranium, cedar, marjoram, myrrh, palmorosis, patchouli, guaiac tree, maracana chamomile and tea tree with a certain content of components [Patent 2416425 Russian Federation IPC A61K 36/899, 36/72, 36/53, 36/185, 36/28, 36/15, 17/02 Antidecubital oil / Krylova V.L.; applicant and patent holder Krylova V.L. - No. 2010107285/15, Appl. 02/27/2010, publ. 04/20/2011, Bull. #11]

Known agent with anti-inflammatory, antiparasitic, antifungal and anti-burn action based on fir oil, characterized in that it contains vegetable, or linseed, or cedar oil, 10-20% alcohol tincture of propolis and fir oil, the rest [Patent 2192273, Russian Federation IPC A61K 35/78, 35/64 Means "natural shield" with anti-inflammatory, antiparasitic, antifungal and anti-burn action / Chukalov A.A. applicant and patentee Chukalov A.A. No. 2001134723/14, Appl. 12/24/2001 publ. November 10, 2002, Bull. No. 31].

Being biologically active substances, essential oils have a healing effect on the affected areas of the skin when they are used directly in the treatment and prevention of bedsores and burns, respectively. However, the direct effect of these substances on the affected areas is short-term and requires constant physical and mechanical action on these areas, which creates inconvenience in their use and reduces the expected therapeutic effect.

To prolong the therapeutic effect of oils, they are enclosed in micro- and nanocapsules consisting of surfactants and polyelectrolytes.

Known composition for the treatment of textile material intended for direct contact with damaged human skin. The composition is a nano- and microemulsion containing nanocapsules and microcapsules, with encapsulated biologically active substances included in them. Natural sea buckthorn oil is used as a biologically active substance, a nano- and microemulsion is prepared as follows: water is poured into the impregnation bath, then anionic surfactant, nonionic surfactant, natural sea buckthorn oil are sequentially introduced, with constant stirring at a rotation speed of 900-6500 rpm and gradually bring the volume of the nano- and microemulsion to a predetermined volume with water, everything is thoroughly mixed for 1-5 minutes until a homogeneous nano- and microemulsion is obtained, after which a cationic polyelectrolyte is added to the obtained nano- and microemulsion and again everything is thoroughly mixed for 0, 5-1 min.

All these drugs have an oil-in-water emulsion composition, which limits the possible options for the biologically active substances used by the hydrophobic nature of the drugs, the therapeutic efficacy of which is inferior to hydrophilic substances [Volodina, T.A. Possibilities of using phytocompositions and generic drugs for the development of external soft medicinal forms of reparative action / Т.А. Volodina, Yu.Yu. Zhidkova, A.V. Mayorova // Fundamental research. - 2011. - No. 11-1. - S. 198-201; Obolensky, V.N. Chronic wound: a review of modern methods of treatment / V.N. Obolensky // RMJ. - 2013. - No. 5. - S. 282.]. In addition, the disadvantage of using essential oils, even encapsulated ones, are possible allergic reactions and irritation of the skin, which aggravate the patient's painful condition.

Known nanoemulsion with biologically active substances [Patent 2362544 Russian Federation IPC A61K 9/10, 9/107 Nanoemulsion with biologically active substances / Pozharitskaya O.N., Karlina M.V., Shikov A.N., Makarov V.G., Makarova M.N.; applicant and patentee Closed Joint Stock Company "St. Petersburg Institute of Pharmacy", No. 2007113157/15, dec. 04/09/2007 publ. 27.02.2010 Bull. No. 21], which refers to the pharmaceutical, food industry and cosmetology, namely to the field of creating nanoemulsion systems used as carriers of active substances in pharmaceutical compositions, as well as in the production of food and cosmetic products. Transparent or slightly opalescent water-in-oil nanoemulsion for oral, transdermal use, for use in ophthalmic practice, with biologically active compounds is characterized in that it contains 35-80% hydrophobic phase, 17-43% surfactant, 3-7% co-solvent and 1-15% of the aqueous phase, mixtures of mono-, di- and triglycerides with mono- and di-esters of saturated and unsaturated fatty acids are used as the hydrophobic phase, the surfactant is selected from the group of nonionic surfactants - sorbitans in mixtures with an auxiliary surfactant (from the group of polyhydroxyalkanes or monohydric alcohols). At the same time, biologically active compounds of nanoemulsions are flavonoids, betulin, boswellia extract, vitamins, microelements, etc.

The main disadvantage of this emulsion is the low content of water-soluble biologically active substances in comparison with the hydrophobic phase and auxiliary components.

When conducting a comparative search from the prior art, the closest to the proposed technical solution was not identified.

The technical result achieved by using the present invention is a method for obtaining a highly concentrated nanoemulsion of a biologically active substance, enhancing the therapeutic effect and increasing the duration of exposure to the patient's skin in the area of the affected areas.

This result is achieved by the method of obtaining a nanoemulsion with biologically active substances to provide a long-term therapeutic effect of biologically active substances on the patient's skin in the area of the affected areas in the treatment of bedsores and burns, according to the invention , 100 ml of a 1% solution of polyelectrolyte I is added to 0.5 g of calcium carbonate ( 10 g/l), stirred for 20 minutes at 20°C, after which the solution is centrifuged at a speed of 2000-4000 rpm for 10-20 minutes, the obtained particles are washed three times with water at a temperature of 20°C, 100 ml of 1 % solution of polyelectrolyte II (10 g/l), stirred for 20 minutes at 20°C, a solution of biologically active substance (5-15 g/l) is added, an equivalent amount of ethyl alcohol is added and stirred for 20 minutes; in this case, a preparation based on the fungus strain Pleurotus ostreatus VKPM F-819 is used as a biologically active substance, chitosan is used as polyelectrolyte I, xanthan gum or guar gum or sodium alginate is used as polyelectrolyte II.

The achievement of the claimed technical result in the proposed method is due to the use as a biologically active substance of an effective drug with polyfunctional medical and biological activity that affects tissue metabolism. The drug was obtained as a result of liquid-phase deep cultivation of the fungus Pleurotus ostreatus VKPM F-819, followed by separation of the mycelium and culture fluid and isolation of biologically active substances from the mycelium in the form of a condensed extract with antimicrobial activity. The extract is the pharmacological substance of the drug and is enriched with the active principle of the sterol derivative 4-hydroxy-17R-methylincisterol with a molecular weight of 332.2452 daltons, polysaccharide β 1-3 glucan and dihydroquercetin with a molecular weight of 304.26 daltons in the following mass ratio: (1) :(2):(3):(4)as(79.0-158.0):(1.0-2.0):(10.0-20.0):(10.0-20, 0) [Patent 2487930 Russian Federation IPC C12N 1/14, A61K 36/06 A drug with multifunctional medical and biological activity that affects tissue metabolism, based on the strain of the fungus Pleurotus ostreatus VKPM F-819 / Gerasimenya V.P., Zakharov S. V., Kiryanov G.I., Polyakov V.Yu., Applicant and patentee Gerasimenya V.P., No. 2012125336/10, Appl. 06/19/2012, publ. 20.07.2013 Bull. No. 20].

The drug has bactericidal and wound-healing properties, when applied to the damaged cover, it restores blood circulation at the site of injury, provokes the rejection of necrotic masses (burns or bedsores), provides restoration of the skin in the area of the burn or bedsore and the fastest healing of the cleaned wound [Milevich T.I. Oyster mushroom mycelium - the basis of biotechnological therapeutic and prophylactic drugs of a new generation: abstracts / T.I. Milevich, A.D. Naumov, V.P. Gerasimenya, S.V. Zakharov, K.Z. Gumargalieva, L.A. entrepreneurial tool. Offers and commercialization technologies”, Gomel, June 21-22. - 2011. - S. 77-79].

For prolonged release, the drug is encapsulated in a shell based on templates, which are spherical colloidal particles of calcium carbonate CaCO ₃ . Particles are obtained by mixing solutions of calcium chloride CaCl ₂ and soda (or sodium carbonate) Na ₂ CO ₃ with a concentration of 0.33 M.

CaCl ₂ + Na ₂ CO ₃ \u003d CaCO ₃ + 2NaCl

During stirring, an amorphous precipitate of calcium carbonate formed. After completion of the process, the CaCO ₃ templates were washed from Na ⁺ and Cl ^- ions with distilled water and evaporated.

To form the polyelectrolyte shell, biodegradable polyelectrolytes were used: chitosan (I), xanthan gum (II), guar gum (III), and sodium alginate (IV).

To deposit polyelectrolyte layers on particles, we chose the method of polyion assembly, which was carried out by successive treatment with oppositely charged polyelectrolytes. Calcium carbonate nuclei have a negative surface charge; therefore, a positive polyelectrolyte chitosan, which forms a positively charged polycation, was applied as the first layer.

The same procedure was then carried out using a negative polyelectrolyte solution (xanthan gum, guar gum, sodium alginate).

To open the shell of the microcapsule and the penetration of the biologically active substance through the layers of polyelectrolytes deep into the microcapsules, ethyl alcohol was used [GOST 5962-2013].

The invention is carried out as follows

Example 1

To 0.5 g of calcium carbonate, 100 ml of a 1% solution of chitosan is added in an amount of 10 g/l. The resulting mixture is stirred for 20 minutes at 20°C and centrifuged for 20 minutes. at a speed of 2000 rpm. The obtained particles are washed three times with water at a temperature of 20°C, 100 ml of a 1% solution of xanthan gum (10 g/l) are added, stirred at 20°C for 20 minutes. After that, a solution of the VKPM F-819 preparation (5 g/l) and an equivalent amount of ethyl alcohol are introduced into the resulting emulsion. The mixture is stirred for 20 minutes to form microcapsules.

Example 2

To 0.5 g of calcium carbonate, 100 ml of a 1% solution of chitosan is added in an amount of 10 g/l. The resulting mixture is stirred for 20 minutes at 20°C and centrifuged for 15 minutes. at a speed of 3000 rpm. The obtained particles are washed three times with water at a temperature of 20°C, 100 ml of a 1% solution of guar gum (10 g/l) are added, stirred at 20°C for 20 minutes. After that, a solution of the VKPM F-819 preparation (10 g/l) and an equivalent amount of ethyl alcohol are introduced into the resulting emulsion. The mixture is stirred for 20 minutes to form microcapsules.

Example 3

To 0.5 g of calcium carbonate is added 100 ml of a 1% solution of chitosan in an amount of 10 g/l. The resulting mixture is stirred for 20 minutes at 20°C and centrifuged for 10 minutes. at a speed of 4000 rpm. The obtained particles are washed three times with water at a temperature of 20°C, 100 ml of a 1% sodium alginate solution (10 g/l) are added, stirred at 20°C for 20 minutes. After that, a solution of the VKPM F-819 preparation (15 g/l) and an equivalent amount of ethyl alcohol are introduced into the resulting emulsion. The mixture is stirred for 20 minutes to form microcapsules.

Claims (1)

A method for producing a nanoemulsion with biologically active substances to provide a long-term therapeutic effect of biologically active substances on the patient's skin in the area of the affected areas in the treatment of bedsores and burns, characterized in that 100 ml of a 1% solution of polyelectrolyte I 10 g are added to 0.5 g of calcium carbonate /l, stirred for 20 min at 20°C, after which the solution is centrifuged at a speed of 2000-4000 rpm for 10-20 min, the obtained particles are washed three times with water at a temperature of 20°C, 100 ml of 1% a solution of polyelectrolyte II 10 g/l, stirred for 20 minutes at 20°C, a solution of biologically active substance 5-15 g/l is introduced, an equivalent amount of ethanol is added and stirred for 20 minutes; at the same time, a preparation based on the fungus strain Pleurotus ostreatus VKPM F-819 is used as a biologically active substance, chitosan is used as polyelectrolyte I, xanthan gum, or guar gum, or sodium alginate is used as polyelectrolyte II.