WO2021167481A1

WO2021167481A1 - Micellized solution of fullerenes, and method for the production and use thereof

Info

Publication number: WO2021167481A1
Application number: PCT/RU2020/000082
Authority: WO
Inventors: Екатерина Михайловна ВОЛОЧАЕВА; Екатерина Михайловна ФЕДОРОВА; Анатолий Владимирович САМОЙЛОВ; Даниил Александрович КОШЕЛЕВ
Original assignee: Акционерное Общество "Акванова Рус"
Priority date: 2020-02-19
Filing date: 2020-02-19
Publication date: 2021-08-26

Abstract

The invention relates to micellized solutions of fullerenes, which are used in cosmetic and pharmaceutical products and in biologically active dietary supplements. A method of producing a micellized solution of fullerenes comprises heating a surfactant to a temperature of not more than 45-75°C while mixing, adding to the resulting solution an oil solution of fullerenes, and subsequently heating the resulting mixture to a temperature of not more than 70-95°C while mixing until a transparent solution forms, wherein the surfactant content in the solution is at least 85 wt% and the content of oil solution of fullerenes is not more than 15 wt%. The invention makes it possible to produce a stable micellized solution of fullerenes that simultaneously has both water- and fat-soluble properties, has improved bioavailability, is capable of maintaining transparency when diluted with water, and can be used as an active ingredient in the manufacture of biologically active dietary supplements, cosmetics and agents for skin care or regeneration.

Description

MICELATED FULLERENE SOLUTION, METHOD FOR ITS PREPARATION AND

APPLICATION

The invention relates to the field of chemistry, in particular to micellized solutions of fullerenes, which can be used in cosmetic and pharmaceutical products and biologically active additives (BAA).

State of the art

The most common method for obtaining fullerenes is their production from soot obtained by burning graphite electrodes in an electric arc under certain conditions. The resulting soot can contain up to 10% of a mixture of C60 and C70 fullerenes, which are then recovered in pure form by dissolving the soot in organic solvents, filtration, centrifugation, and evaporation. Highly concentrated fullerene mixtures obtained in this way have a high cost due to the need for concentration and subsequent purification from residual solvents.

For use in dietary supplements or cosmetics, the fullerenes obtained in this way must be re-converted into the form of a solution, because their use in solid form is impractical. The residual content of solvents in fullerenes can have a negative effect on the human body. In this regard, in the production of dietary supplements, cosmetic and pharmacological agents, an oily solution is most often used, which is obtained by dissolving fullerene-containing soot in vegetable oil (sunflower, olive, etc.) in an amount of up to 10%, followed by filtration. The cost of this form is much lower due to the elimination of the stages of cleaning from the solvent.

The disadvantage of using this form is the low level of bioavailability of fullerenes. To achieve the desired effect, it is required to increase the recommended dosage of the drug for the consumer, which entails an increase in consumer costs for the purchase of the product. In addition, excessive consumption of oils may be contraindicated in some categories of consumers.

For the production of cosmetics and medicines and dietary supplements, the most preferable is the use of fullerenes in a water-soluble form, since this form has a higher bioavailability compared to oily forms.

There are several ways to obtain water-soluble forms, for example, by attaching amino acid residues to the surface of the fullerene molecule. Arginine or proline residues or derivatives are preferably used. However, such solutions are sensitive to the effects of microorganisms, and the technology for their preparation is rather complicated. The factors described significantly limit the use of such forms of fullerene solutions in the production of cosmetic and pharmaceutical products.

From the international application WO2019052471 a method is known for obtaining a water-soluble form of a solution of fullerenes (emulsion of fullerenes) by mixing an emulsifier and an oil solution of fullerenes preheated to 55-70 ° C and then dispersing the resulting mixture at a speed of 3000-7000 rpm for 8-15 minutes. When this oily solution of fullerenes is obtained from powder of fullerenes by extraction with a solvent of polyethylene glycol-15-hydroxystearate. The emulsion form of fullerenes obtained in this way is a dispersed system consisting of microscopic droplets of a dispersed phase (in this case, fullerene oil), distributed in a dispersed medium (aqueous phase) with the addition of an emulsifier.

A significant disadvantage of the known method is that polyethylene glycol-15-hydroxystearate is not included in the list substances permitted for use in the food industry, and the micellized solution obtained in this way cannot be used in the production of biologically active additives. The resulting solution of fullerenes is not suitable for use in its pure form; it must be diluted with water. Another significant disadvantage of the known method is the duration of its implementation - from 48 to 80 hours.

From the RF patent DO2303036, a method is known for preparing a solubilizate based on ascorbic acid and an emulsifier with a hydrophilic-lipophilic balance (HLB) value from 9 to 18, in which 10 wt.% Of ascorbic acid is dissolved in 10 wt.% Distilled water, into a solution with stirring and heating up to about 80 ° C, add 80 wt.% polysorbate 80, the resulting solution is stirred until transparency and homogeneity. In another embodiment, 10% by weight of ascorbic acid is dissolved in 10% by weight of distilled water, 70% by weight of polysorbate 80 is added to the resulting aqueous solution together with 10% by weight of light vegetable oil, such as, for example, thistle or linseed oil, and then the solution is heated to at least 60 ° C and stirred until transparent and homogeneous. Ascorbic acid in the solubilization product is present in the form of micelles. Microscopic examination of the solubilization product containing 10% ascorbic acid in a dilution of 1: 1000 revealed micelles with a diameter of approximately 100 nm. The solubilizate can be used as an additive to food or cosmetics, pharmaceuticals or nutrient solutions for the cultivation of cell and bacterial cultures and algae.

The known technical solutions do not make it possible to obtain a fullerene-based product with improved bioavailability and good stability. Disclosure of invention

The aim of the claimed invention is to obtain a stable micellized solution of fullerenes, which has both water and fat-soluble properties, as well as improved bioavailability, is able to maintain transparency when diluted with water and can be used as an active ingredient in the production of biologically active additives (BAA), cosmetics or products for the care or restoration of the skin.

The micellized solution of fullerenes according to the claimed invention has a number of advantages over the conventionally used forms of fullerenes.

First, the micellized form of fullerenes has an increased bioavailability compared to the oil solution, from which fullerenes are absorbed in the body in insignificant amounts.

Secondly, the micellized form makes it possible to obtain aqueous solutions that can be added to dietary supplements or cosmetic products if the use of an oil solution is impossible or difficult.

Thirdly, the micellized form of fullerenes is resistant to the action of microorganisms and is capable of retaining its structure and properties for a long time.

Fourthly, the composition of the micellized form of fullerenes includes surfactants (surfactants) approved for use in the food, cosmetic and pharmaceutical industries. Due to this, one form of micellized fullerenes can be used in different types of products, while no additional modification or modification of the micellized form is required, depending on the required field of its application.

The main components of the micellized solution, according to the claimed invention, are an oil solution of fullerenes and a surfactant (surfactant) with a hydrophilic-lipophilic balance (HLB) of 9 or more.

To obtain a micellized solution, an oily solution of fullerenes is added to a surfactant (with an HLB value of 9 or more) preheated to a temperature of 45-75 ° C with simultaneous stirring. Heating the surfactant to a temperature of 45-75 ° C is necessary to facilitate subsequent mixing. This temperature is the optimum temperature at which the surfactant has a sufficiently fluid consistency to ensure uniform distribution of the fullerene oil solution inside it during subsequent stirring. A higher temperature only increases the overall process time, but does not affect the mixing process. The resulting mixture is heated, without stopping stirring, to a temperature of 70-95 ° C. The specified temperature range is optimal for the formation of micelles. At higher temperatures, micelles may not form.

The mixture is kept until a clear solution is formed, while the surfactant is taken in an amount of at least 85 May. %. The total mixing time is no more than 3 hours. This gives micelles containing fullerenes inside them, with a shell of the molecules of the surfactant used.

In other words, in the claimed invention, fullerenes are in the form of micelles, i.e. agglomerates consisting of a micellized substance (fullerenes) surrounded by a surfactant, without the addition of water. This form of fullerenes makes it possible to use the micellized solution in its pure form without additional dilution and, accordingly, without the need for additional technological operations.

Fullerenes in micelles are able to easily penetrate the lipid barrier of cells due to the shell of surfactants. At room temperature, a micelle solution is a transparent viscous liquid with a micelle size in the range from 3 to 20 nm.

Due to the nanoscale of micelles, the resulting micelled solution can be diluted with water without turbidity, since the size of micelles formed in the solution is less than the light wavelength, which ensures the stability of the micellized solution and its high bioavailability.

The shelf life of the micellized form of fullerenes is at least 1 year.

An oily solution of fullerenes, preferred for use in the claimed invention, is obtained by dissolving fullerene-containing soot containing C60 and C70 fullerenes in vegetable oil. To obtain an oil solution, the mixture is heated to a temperature of 40-60 ° C and kept under constant stirring for several hours. The content of fullerenes in soot is not more than 10 wt%. Refined deodorized sunflower or olive oil can be used as vegetable oil. The undissolved soot particles are separated from the resulting oil by centrifugation and filtration at room temperature.

Surfactants with an HLB value of 9 or more can be used as surfactants, such as, for example, polysorbate 20, polysorbate 80, polyethylene glycol and its derivatives, esters of sucrose and fatty acids, and mixtures thereof.

In the finished micellized solution, the content of the oil solution is from 5 to 15 wt.% (With a content from 0.01 to 0.1 wt.% Of fullerenes), the rest is the surfactant.

The micellized solution according to the invention can be used either ready-made or in the form of an aqueous solution.

For use in dietary supplements, cosmetics or pharmaceuticals, no additional processing of the micellized solution is required. In one of the options for using a micellated solution of fullerenes, in particular, to obtain a more fluid consistency of the solution, heating the micellized solution to 35 ° C is permissible.

When using a micellized solution in drinks or, if necessary, adding to the final product in the form of an aqueous solution, a micellized solution of fullerenes can be diluted with water and used in the form of an aqueous solution, which does not reduce its bioavailability and effectiveness.

Depending on the field of application of the micellized solution and the purpose of the final product in which it is used, additional active components can be added to the micellized solution, such as, for example, vitamins and vitamin-like substances (flavonoids, choline, coenzyme Q10, lipoic acid, carnitine, etc.) .p.) or plant extracts.

Brief Description of Drawings

FIG. 1 is a graph illustrating particle size distributions in a micellized solution of fullerenes.

The results were obtained by dynamic light scattering. The drawing shows the distribution of particles in solution according to their size. Columns (a) show which particle size scatters light the most during the measurement. Thus, one can judge the quantitative ratio of particles of different sizes in the solution.

Curve (b) is a distribution function and shows the proportion of particles that are smaller or equal in size.

FIG. 2 shows curves illustrating the pharmacokinetics of the content of oil and micellized solutions of C60 fullerenes in whole blood.

FIG. 3 shows curves illustrating the pharmacokinetics of the content of oil and micellized solutions of C60 fullerenes in the lungs. FIG. 4 shows curves illustrating the pharmacokinetics of the content of oil and micellized solutions of C60 fullerenes in the liver.

FIG. 5 shows curves illustrating the pharmacokinetics of the content of oil and micellized solutions of C60 fullerenes in the spleen.

Implementation of the invention

Example 1. Preparation of a micellized solution of fullerenes

To obtain a micellized solution of fullerenes, 95 g of polysorbate 80 is heated to 75 ° C and 5 g of an oil solution of C60 fullerenes with a concentration of 0.5 g / l are added and mixed. The resulting mixture is heated to 95 ° C with constant stirring and stirring is continued until the solution becomes transparent and homogeneous. Thereafter, the micelle solution is cooled to room temperature.

The solution is stable for at least one year.

Example 2. Preparation of a micellized solution of fullerenes

To obtain a micellized solution of fullerenes 85 g of a mixture of polysorbate 20 with sucrose oleate in a ratio of 10: 1 is heated to 45 ° C and 15 g of an oil solution of C70 fullerenes with a concentration of 0.4 g / l is added and stirred. The resulting mixture is heated to 70 ° C with constant stirring and stirring is continued until the solution becomes transparent and homogeneous. Thereafter, the micelle solution is cooled to room temperature.

The solution is stable for at least one year. Example 3. Preparation of a micellized solution of fullerenes

To obtain a micellized solution of fullerenes, 90 g of a mixture of polyethylene glycol is heated to 60 ° C and 10 g of an oil solution of C60 fullerenes with a concentration of 0.5 g / l is added and stirred. The resulting mixture is heated to 80 ° C with constant stirring and stirring is continued until the solution becomes transparent and homogeneous. Thereafter, the micelle solution is cooled to room temperature.

The solution is stable for ... at least one year.

Similar results were achieved when using other surfactants with an HLB value of 9 or more, preferred for the preparation of a micellized solution, according to the claimed invention and at other ratios of components from the claimed ranges.

Example 4. Determination of bioavailability and pharmacodynamics of fullerene solution

The bioavailability and pharmacodynamics of a solution of fullerenes were determined in a mouse model by comparing changes in the content of micellized and oily forms of SbO fullerenes in tissues and organs of mice after intragastric administration.

The dosage of each form of fullerenes was 15 mg / kg of the weight of the animal, which, in terms of the actual content of fullerenes, is 1.5 mg / kg for the oily form of fullerenes and 0.15 mg / kg for the micellized form of fullerenes.

The measurement of the content of fullerenes was carried out by HPLC-MS within 24 hours after the administration of the finished forms to the animals. Samples of blood, lungs, thymus, heart, liver, spleen, kidneys, adrenal glands, brain tissue and testicles were taken for the study.

Figures 2-5 show curves illustrating the pharmacokinetics of the content of oil and micellized solutions of C60 fullerenes in the examined tissues and organs.

It follows from the results obtained that fullerenes SbO in oily and micellized forms accumulate in tissues in an identical manner. The detectable amounts of fullerenes are determined in the liver, lungs, kidneys, heart, adrenal glands, thymus and spleen, but are absent in the tissues of the brain and testicles. At the same time, despite the fact that the content of fullerenes in the micellized form is ten times less than in the oily form, their content in the tissues is at the same level or even higher than in the oily solution.

The results obtained confirm that the micellized solution of fullerenes, according to the claimed invention, has an increased bioavailability in comparison with the oil solution of fullerenes, at least tenfold.

Claims

CLAIM

1. A method of obtaining a micellized solution of fullerenes, including heating a surfactant to a temperature of no more than 45-75 ° C with simultaneous stirring, adding an oil solution of fullerenes to the resulting solution and then heating the resulting mixture to a temperature of no more than 70-95 ° With simultaneous stirring until a clear solution is formed, while the surfactant content in the solution is at least 85 wt%, and the content of the oil solution of fullerenes is not more than 15 wt%.

2. A method of obtaining a micellized solution according to claim 1, in which at least one substance from a surfactant with a hydrophilic-lipophilic balance of 9 or more can be used as a surfactant: polysorbate 20, polysorbate 80, polyethylene glycol and its derivatives, sucrose esters and fatty acids, as well as mixtures thereof.

3. A method of obtaining a micellized solution according to claim 1, in which the oil solution of fullerenes is obtained by mixing vegetable oil and fullerene soot, then heating the resulting mixture to 40-60 ° C and then keeping it under constant stirring for several hours.

4. A method for preparing a micellized solution according to claim 3, wherein the fullerene soot contains fullerenes C60 and C70.

5. A method of obtaining a micellized solution according to claim 3, in which the fullerene content in soot is not more than 10 May. %.

6. A method of obtaining a micellized solution according to claim 3, in which sunflower or olive oil can be used as the vegetable oil.

7. A micellized solution of fullerenes obtained by the method of claim 1, wherein said solution comprises a surfactant in an amount of at least 85 wt% and an oily solution of fullerenes.

8. A micellized solution according to claim 7, in which the surfactant has a hydrophilic-lipophilic balance of 9 or more.

9. A micellized solution according to claim 7, wherein the micelle size is in the range of 3-20 nm.

10. A micellized solution according to claim 7, wherein the fullerene content in the oil solution is from 0.01 to 0.1 wt%.

11. The use of a micellized solution according to claim 7 as an active component of biologically active additives (BAA), cosmetics and skin care and / or restoration products.

12. Use according to claim 11, wherein the micellized solution is diluted with water and used in the form of an aqueous solution.

13. Use according to claim 11, wherein the micelated solution can be heated to a temperature not exceeding 35 ° C before use.

14. Use according to claim 11, wherein the micelated solution can be used in conjunction with vitamins, vitamin-like substances and / or plant extracts.