RU2736504C1 - System of transdermal transfer enhancers and cosmetic compositions based thereon - Google Patents

Abstract

FIELD: cosmetology.SUBSTANCE: group of inventions relates to cosmetology. Transdermal transfer enhancers system contains dimethylsulphoxide and pyrrolidone carboxylate of sodium, wherein ratio of components in wt%: dimethylsulphoxide - 0.01-0.05, sodium pyrrolidone carboxylate (NaPCA) - 1-2.5, water - the rest. What is also disclosed is a cosmetic composition.EFFECT: group of inventions enables penetration of biologically active substances through the stratum corneum of the epidermis into the dermis without injuring the skin with preservation of skin moisturisation and fast recovery of the skin barrier after application.12 cl, 3 tbl, 2 ex, 3 dwg

Description

FIELD OF TECHNOLOGY

[001] The present invention relates to the field of cosmetology, in particular to the development of a system of enhancers of transdermal transfer of biologically active substances (BAS). The claimed system of enhancers can be used to create cosmetic compositions containing biologically active substances, or as an independent agent that increases the effectiveness of cosmetics used immediately after using the enhancer system.

TECHNOLOGY LEVEL

[002] More and more people are resorting to minimally invasive and non-invasive cosmetic procedures. Minimally invasive procedures include injections with dermal fillers or biologically active substances, which improve the appearance of the skin without surgery. Since anesthesia is generally not required, most injection procedures pose significantly less risk to the patient than plastic surgery. In addition, cosmetic injections do not require a long rehabilitation period. However, complications such as swelling, bruising, pain, itching, soreness, lines under the skin, and allergic reactions have been reported following cosmetic injections. Injections can also damage blood vessels, resulting in embolism, necrosis, or other significant damage to the skin.

[003] Non-invasive cosmetic procedures involve transdermal (percutaneous) delivery of biologically active substances. The transdermal route of delivery of cosmetic active substances makes it possible to carry out a slow, introduction of biologically active substances and to suspend it if unwanted side effects occur during the cosmetic procedure, for example, allergic reactions. However, the skin has excellent barrier properties, which greatly limits the types of molecules that can be introduced through it.

[004] Structurally, the skin consists of two main layers - a relatively thin outer layer (epidermis) and a thick inner layer (dermis). The epidermis consists of a stratified epithelium, which is characterized by the fact that keratinization of cells constantly occurs in its outer layers. The most superficial stratum corneum of the epidermis consists of corneocytes - anucleated, flat, ordered keratinized cells filled with filamentous keratin. Corneocytes turn into scales, which gradually slough off on the surface of the epidermis, being replaced by new cells originating from deeper layers of the epidermis. The stratum corneum of the epidermis in thick skin consists of 15-20 layers of horny scales, and in thin skin - of 3-4 layers of horny scales [1]. The stratum corneum of the epidermis has a unique structure, which is called "briсk and mortar" ("brick and cement"), where the role of "bricks" is played by corneocytes, and "cement" is the intercellular lipid matrix [2]. Intercellular lipids are represented by ceramides, cholesterol, fatty acids, phospholipids, glycosylceramides, as well as free sphingoid bases and cholesterol sulfate. Intercellular lipids are responsible for the impermeability of the stratum corneum of the epidermis. The dermis is composed of fibrous connective tissue. The dermis contains blood and lymph vessels, nerves, smooth muscle cells, hair follicles, sebaceous and sweat. Sebaceous glands are located throughout the body, except for the skin of the palms and soles. The secretion of the sebaceous glands (sebum), which is produced by special cells - sebocytes, is secreted onto the surface of the epidermis or into the hair funnel. Sebum has bactericidal and fungicidal properties. Sebum is 60% composed of triglycerides, which, under the influence of microbial hydrolases, are converted into free fatty acids, monoglycerides, linoglycerides and glycerol. The rest of the sebum is mainly waxy esters, squalene and cholesterol. Squalene is an intermediate in the biological synthesis of cholesterol. In human sebum up to about 25 years, squalene is about 13% [3]. Sebum prevents the evaporation of endogenous water and, in cold weather, helps to retain heat [1].

[005] According to modern concepts, there are the following ways of penetration of exogenous substances through the skin: transdermal (through the stratum corneum of the epidermis), transfollicular (through the hair follicles) and transglandular (through the ducts of the sebaceous and sweat glands). At the same time, from 0.01% to 0.1% of exogenous substances penetrating through the stratum corneum penetrates through the hair follicles, ducts of the sebaceous and sweat glands [4] Thus, the barrier properties of the skin are mainly determined by the barrier properties of the stratum corneum of the epidermis, which protects the underlying tissues from infections, dehydration, from aggressive chemical and physical effects [5] The permeability of the stratum corneum is mainly due to the permeability of the lipid matrix, which fills the gaps between corneocytes, but it is possible for exogenous substances to pass directly through corneocytes.

[006] Effective penetration of exogenous substances into the deep layers of the skin along the concentration gradient is possible only for molecules that have an affinity for both the hydrophobic stratum corneum and the underlying hydrophilic layers of the epidermis and dermis. That is, the molecules must be neutral. A positive or negative charge of a molecule can slow down its movement through a hydrophobic medium. For example, the efficiency of penetration of charged substances through the stratum corneum of the skin is two orders of magnitude lower than that of uncharged substances [6]. In addition, the penetrating ability of an exogenous substance depends on the molecular weight of the molecules.

[007] Permeability is also influenced by the localization of the skin area, the degree of its hydration, the thickness of the stratum corneum, the presence or absence of a lipid lubricant and its qualitative composition. Without special transfer systems, only small low-polarity molecules - alcohols, ketones, etc. can penetrate into healthy, intact skin. Physical and chemical approaches are mainly used to overcome the barrier properties of the stratum corneum.

[008] In a physical approach to the transdermal delivery of biologically active substances, in order to create reversible physical changes within the stratum corneum, external forces are used: electric current (iontophoresis), high-voltage electrical impulses (electrophoresis / electroporation) and ultrasound (sonophoresis). This modification of the stratum corneum allows transdermal delivery of large charged molecules that cannot pass through the stratum corneum due to passive diffusion. The main advantages of this approach are: rapid penetration of biologically active substances and, accordingly, the short time required for biologically active substances to reach the target, as well as the ability to control delivery by varying the strength and duration of exposure. The disadvantages of the physical approach are the need to use specific equipment, possible unpleasant sensations from the physical impact used and contraindications to the use of certain types of impact in a number of diseases.

[009] One of the chemical approaches is to deliver the biologically active substances in an encapsulated form, for example, inside liposomes capable of passing through the stratum corneum of the skin with the subsequent release of the biologically active substances. However, the use of liposomes in cosmetics and pharmaceuticals is limited due to their low thermodynamic stability and low capacity [7].

[0010] The main disadvantage of compositions containing liposomes and microemulsion solutions as carriers of biologically active substances is their delamination upon introduction of an aqueous or non-polar BAS solution, which causes technological problems in the production of cosmetics and pharmaceuticals based on them.

[0011] Another popular chemical approach to the transdermal delivery of biologically active substances is the use in the composition of a cosmetic or drug of additional chemicals - transdermal transfer enhancers, which act on the membranes of corneocytes or modify the extracellular lipid matrix of the stratum corneum, thereby increasing the permeability of the skin barrier.

[0012] In the last two decades, a large number of compounds have been studied in terms of their effectiveness for increasing the rate and intensity of penetration of biologically active substances through the skin. A wide range of substances are used as enhancers, including polyhydric alcohols, fatty acids and their esters, surfactants, and terpenes. Classical examples of enhancers are proton acceptor solvents, in particular dimethyl sulfoxide (DMSO) and dimethylacetamide (DMAA). In addition, 2-pyrrolidone, N, N-diethyl-m-toluamide (DETA), 1-dodecylazacycloheptan-2-one, N, N-dimethylformamide, N-methyl-2-pyrrolidone, and calcium thioglycollate have been described as effective enhancers.

[0013] In patents US4980378 (published 12.25.1990) and US5082866 (published 01.21.1992) a group of biodegradable is disclosed, which are N, N-disubstituted aminoacetates. These biodegradable transdermal transfer enhancers do not induce a reaction and / or irritation on the skin at the site of application. The disadvantages of using N, N-disubstituted aminoacetates include the limited number of biologically active substances with which they can be used.

[0014] Some carriers are nonionic surfactants consisting of a polar group and a high molecular weight non-polar moiety. These compounds include N-dodecylcaprolactam-2, isopropyl myristate, oleic acid, analogs of N-dodecylcaprolactam-2. All these substances interact with the lipid structure of the epidermis, destroying its orderliness. As a result, the resistance of the skin to the flow of biologically active substances decreases.

[0015] The most commonly used chemical enhancer is DMSO, a representative of the class of bipolar aprotic substances. DMSO is less toxic than other members of this group, such as dimethylformamide, dimethylacetamide, N-methyl-2-pyrrolidone, etc. DMSO demonstrates low toxicity to animals, plants or aquatic organisms, is not a carcinogen [8] and does not exhibit mutagenic properties in standard tests [9]. No genotoxicity has been reported following the use of DMSO as a drug, and no similar effects have been reported in workers during 40 years of industrial production [10].

[0016] It should be noted that DMSO has anti-inflammatory, decongestant and local analgesic effects [11].

[0017] DMSO is able to penetrate biological membranes, facilitating better and deeper penetration into the skin of biologically active substances used simultaneously with it. Rama et al. describes the creation and study of in vivo and in vitro transdermal pharmaceutical compositions of methotrexate based on TWIN-80, Span-80, DMSO and isopropyl myristate. There is a high penetrating ability of DMSO in comparison with other agents [12].

[0018] Numerous cosmetic formulations are known using DMSO, for example, application WO2017205659 (published: 05/25/2017), patent EA019760B1 (published: 10/31/2008), application WO2011000210 (published: 06/24/2010). In all of these formulations, DMSO is used as a transdermal transfer enhancer.

[0019] However, for all its advantages, DMSO has a number of side effects. For example, at a concentration of 25-100%, DMSO destroys cell membranes. With topical application of DMSO at a concentration of 45.5% to 90%, redness, itching or burning are observed at the site of application, as well as the appearance of an unpleasant garlic odor from the mouth, which occurs due to the fact that in the human body DMSO is metabolized to dimethyl sulfone and dimethyl sulfide, which excreted through the lungs [13–21].

[0020] Ethanol is another known enhancer of transdermal transfer of lipophilic biologically active substances. Known composition RU2107515 comprising a water-ethanol solvent containing 15-75% ethanol by volume. Over the entire specified concentration range, ethanol provides a marked improvement in transdermal penetration. However, ethyl alcohol causes significant skin dehydration.

[0021] Dehydration weakens the protective function of the skin, promotes the development of immune disorders and the formation of allergic inflammation [22]. Numerous structures are involved in maintaining the water balance of the skin that regulate the rate of transepidermal water loss (TEWL) and carry out the flow of water from the dermis into the epidermis. In the epidermis, the control of the water balance is carried out by the components of the stratum corneum - a natural moisturizing factor, skin lipids, sebum and keratin [23, 24].

[0022] The natural moisturizing factor (NMF) is a complex of organic molecules on the surface of corneocytes that has the ability to bind water. NMF contains free amino acids, NaPCA, urea, ammonia, creatinine, magnesium, potassium, calcium, sodium, lactic and citric acids, chloride and phosphate ions. NMF binds approximately one third of the water molecules contained in the stratum corneum of the epidermis. In addition, NMF maintains normal exfoliation of dead cells through the participation of hydrolytic enzymes. The imbalance of the NMF components reduces the ability of the epidermis to retain moisture [25–27].

[0023] 2-Pyrrolidone-5-carboxylic acid (PCA) is a component of NMF and, along with sodium lactate, constitutes the most hygroscopic fraction of the stratum corneum of the epidermis [27, 28]. The overwhelming amount of PCA in the stratum corneum at a pH of about 5 exists in the form of salts, including the sodium salt. Thus, the sodium salt of 2-pyrrolidone-5-carboxylic acid, or sodium pyrrolidone carboxylate (NaPCA), is a natural constituent of the human epidermis and is often used in cosmetics as a moisturizing additive. For example, in applications and patents CN110051614 (published: 05/27/2019), CN107837205 (published: 11/28/2017), CN108578277 (published: 02/13/2016), CN104546654 (published: 10/11/2013), RU2444980 (published: 03/20/2012) describes cosmetics containing NaPCA as a hygroscopic substance, which, at small molecular sizes, is capable of binding large amounts of water. For 2019, NaPCA is used in 1289 cosmetic products [29].

[0024] A wide range of various substances are used as BAS in cosmetic compositions, including peptides, amino acids, nucleic acids, hyaluronic acids, saccharides, ceramides, vitamins, minerals and many others. However, most of the biologically active substances do not independently penetrate through the stratum corneum of the epidermis into the deeper layers of the skin, where they should exert their effect. In connection with the development of non-invasive cosmetology, there is a need to develop a system of transdermal transfer enhancers that will allow biologically active substances to successfully overcome the skin barrier.

TERMS AND ABBREVIATIONS

[0025] EGF - epidermal growth factor

[0026] MgPCA - magnesium 2-pyrrolidone carboxylate

[0027] MMP - Matrix Metalloproteinase

[0028] NaPCA - sodium 2-pyrrolidone carboxylate

[0029] NMF (natural moisturizing factor) - natural moisturizing factor

[0030] PCA - 2-pyrrolidone-5-carboxylic acid

[0031] TGF-β - transforming growth factor β

[0032] VEGF - vascular endothelial growth factor

[0033] ROS - reactive oxygen species

[0034] BAS - biologically active substances

[0035] DMSO - dimethyl sulfoxide

[0036] kDa - kiloDalton

[0037] UV - ultraviolet

[0038] Vasculoendothelial growth factor (VEGF, Vascular endothelial peptide, Vascular endothelial growth factor) is a series of signal peptides that result from the alternative splicing of messenger RNA (mRNA) of a single gene containing 8 exons. VEGF is expressed by various cell types under oxygen-deficient conditions. VEGF serves as part of the system responsible for restoring oxygen supply to tissues.

[0039] Vitamin A is a series of retinoids including retinol, tretinoin, adapalene, tazarotene retinaldehyde and retinyl esters.

[0040] Vitamin B - group of water soluble vitamins include vitamin B1 (thiamine, aneurine), vitamin B2 (riboflavin, lactoflavin), vitamin B3 (niacin, niacin), Vitamin B5 (pantothenic acid), vitamin B6 (pyridoxine, pyridoxal , pyridoxamine, pirivitol), vitamin B7 (biotin, vitamin H), vitamin B9 (folic acid), vitamin B12 (cobalamin, cyanocobalamin).

[0041] Vitamin C (L-ascorbic acid) is a biologically active isomer of ascorbic acid that is necessary for the normal functioning of connective and bone tissue.

[0042] Vitamin E (tocopherol) is a group of naturally occurring fat-soluble tocol derivatives. The most active form of vitamin E is α-tocopherol.

[0043] Vitamin F - is a complex of polyunsaturated fatty acids belonging to the classes of omega-3-unsaturated fatty acids and omega-6-unsaturated fatty acids. The complex includes linoleic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid. The name "vitamin F" is arbitrary, since from the biochemical and pharmacological point of view, the specified complex of polyunsaturated fatty acids is not a vitamin, and it is isolated into a special group of biologically active substances - essential fatty acids.

[0044] Vitamin K is the group name for a number of fat-soluble, hydrophobic vitamins derived from 2-methyl-1,4-naphthoquinone. Vitamin K1 isolated from alfalfa and K2 isolated from rotting fishmeal have been found in nature. In addition to natural vitamins K, a number of synthetic naphthoquinone derivatives are currently known: vitamin K3 (2-methyl-1,4-naphthoquinone), vitamin K4 (2-methyl-1,4-naphthohydroquinone), vitamin K5 (2- methyl 4-amino-1-naphthohydroquinone), vitamin K6 (2-methyl-1,4-diaminonaphthoquinone), vitamin K7 (3-methyl-4-amino-1-naphthohydroquinone).

[0045] Tripeptide-10 citrulline (Lys-a-Asp-Ile-Citrulline, Decorinyl, Decorinyl ^TM ) is a synthetic tetrapeptide, an analogue of the decorin protein produced by human skin.

[0046] Carnosine (L-carnosine, β-Ala-His peptide) is a low molecular weight water-soluble dipeptide consisting of β-alanine and histidine amino acid residues. Found in high concentrations in the muscles and brain tissues of mammals, including humans.

[0047] Palmitoyl pentapeptide-3 (pal-KTTKS, Matrixyl, Matrixyl ^®) - lipopeptide C ₁₆ -KTTKS, prepared by proteolytic hydrolysis of collagen type I and stabilized due palmitoylation.

[0048] Palmitoyl tripeptide-38 (pal-Lys-Met-Lys peptide) is a signal peptide consisting of two lysine and one methionine residues, stabilized by palmitation.

[0049] Peptide Ala-Glu-Asp-Gly (AGAG peptide, epithalon) is a tetrapeptide synthesized based on the analysis of the amino acid composition of a complex peptide preparation of epithalamin isolated from the pineal gland of the animal brain.

[0050] Peptide Arg-Lys-Asp-Val-Tyr (RKDVY peptide) is a pentapeptide corresponding to a region of the amino acid sequence of the thymopoietin protein produced by the thymus. The sequence of amino acid residues Arg-Lys-Asp-Val-Tyr (RKDVY) corresponds to positions 32-36 in the peptide chain of thymopoietin.

[0051] Peptide Cu-GHK (Copper tripeptide, peptide GHK-Cu, peptide Cu-Gly-L-His-L-Lys) is a natural complex of monovalent copper and a tripeptide consisting of glycine, histidine and lysine.

[0052] Peptide Glu-Trp (peptide EW) is a synthetic dipeptide identical to a natural compound isolated by chromatography from thymus extract.

[0053] Peptide Lys-Glu (peptide KE) is a synthetic dipeptide consisting of lysine and glutamic acid residues, one of the representatives of peptide thymomimetics, which was found in the thymus polypeptide complex of animals.

[0054] Peptide Lys-Asp-Glu (KDE peptide) is a vasoactive tripeptide composed of lysine, aspartic acid and glutamic acid residues.

[0055] Free amino acid is an amino acid that is not part of the peptide.

[0056] Squalane is a triterpene hydrocarbon belonging to the group of carotenoids. Squalane is a stabilized form of squalene obtained from a hydrogenation process. Squalane is more resistant to oxidation and high temperatures than squalene. Belongs to the group of carotenoids.

[0057] Epidermal growth factor (EGF) is a globular protein of about 6 kDa, consisting of 53 amino acid residues. Also, the term "EGF" in the text of this application means synthetic peptides, the amino acid sequences of which correspond to a part of the amino acid sequence of EGF.

SUMMARY OF THE INVENTION

[0058] The objective of the present invention is the non-invasive delivery of biologically active substances in cosmetic compositions into the deep layers of the skin, including the dermis.

[0059] This problem is solved by the claimed invention by achieving such a technical result as ensuring the penetration of biologically active substances through the stratum corneum of the epidermis into the dermis without injuring the skin, rapid restoration of the skin barrier after applying the claimed invention, maintaining skin moisture.

[0060] The claimed technical result is achieved due to the composition of the system of transdermal transfer enhancers, which contains the components listed below in the following ratio wt. %:

Dimethyl sulfoxide (DMSO) - 0.01-0.05,

Sodium pyrrolidone carboxylate (NaPCA) - 1-2.5,

Water - 97.45-98.99

[0061] The claimed system of enhancers can be used independently, immediately before applying the cosmetic. In this case, the claimed system of enhancers increases the effectiveness of this cosmetic product, increasing the permeability of the skin to biologically active substances contained in the cosmetic product.

[0062] DMSO and NaPCA in the claimed system of enhancers serve to ensure the penetration of biologically active substances through the stratum corneum of the epidermis. At the same time, NaPCa additionally helps to maintain skin hydration. In addition, NaPCa acts as a preservative due to its antimicrobial properties.

[0063] In one embodiment, the claimed system of enhancers contains squalane, which serves to prevent dehydration of the skin, and also facilitates the penetration of certain biologically active substances through the stratum corneum of the epidermis. Squalane additionally has anti-inflammatory properties and helps prevent UV damage to the skin. The squalane content in the claimed enhancer system is 2-5%. In this case, the amount of water in the claimed enhancer system is reduced by that amount by weight. %, which corresponds to the content of squalane.

[0064] The claimed system of enhancers can be included in a cosmetic composition containing a BAS. In this case, the cosmetic composition has the following composition in wt. %:

Dimethyl sulfoxide (DMSO) - 0.01-0.05,

Sodium pyrrolidone carboxylate (NaPCA) - 1-2.5,

BAV - 4-10,

Water - 87.45-94.99.

[0065] In this case, the BAS in the claimed cosmetic composition include at least one peptide, and may also include a wide range of substances, including free amino acids and their derivatives, vitamins, polyunsaturated fatty acids, magnesium and silicon.

[0066] The peptide in the claimed cosmetic composition can be selected from the following: Ala-Glu-Asp-Gly peptide, Lys-Asp-Glu peptide, Lys-Glu peptide, Arg-Lys-Asp-Val-Tyr peptide, Glu- peptide Trp, tripeptide-10 citrulline, carnosine, palmitoyl pentapeptide-3, Cu-GHK peptide, palmitoyl tripeptide-38, vascular endothelial peptide (VEGF), epidermal growth factor (EGF). Moreover, the cosmetic composition may include both one peptide and a combination of peptides, while the total content of peptides in the claimed cosmetic composition does not exceed 0.01-1% by weight.

[0067] Peptides in the claimed cosmetic composition slow down age-related changes in the skin, help smooth wrinkles, reduce age spots, increase moisture, elasticity and smoothness of the skin. Peptides protect skin cells from the negative effects of UV, radiation and toxic-chemical factors, have antimutagenic and anticarcinogenic effects. In addition, peptides have an anti-inflammatory, wound healing, antiseptic effect, thereby improving the condition of the skin in dermatitis, erythema, infectious and inflammatory diseases.

[0068] Free amino acids and their derivatives in the claimed cosmetic composition are involved in maintaining skin moisture, firmness and elasticity. Free amino acids and their derivatives can be represented by one or more variants selected from the range: glycine, alanine, valine, isoleucine, leucine, methionine, tyrosine, serine, hydroxyproline, proline, threonine, cysteine, aspartic acid, asparagine, glutamic acid, glutamine , histidine, lysine, arginine, phenylalanine, tryptophan, ectoine. Moreover, the total content of free amino acids and their derivatives in the claimed cosmetic composition does not exceed 5% wt.

[0069] The vitamins in the claimed cosmetic composition reduce wrinkles, increase skin elasticity, help maintain skin moisture, and protect the skin from sunburn. Vitamins regulate the work of the sebaceous glands, have an anti-inflammatory effect and promote wound healing for minor injuries, thereby improving the condition of the skin in diseases such as acne, dermatitis, psoriasis, eczema. Vitamins can be represented by one or more options selected from the series: vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, vitamin E, vitamin C and vitamin K. the total content of vitamins in the claimed cosmetic composition does not exceed 2-5% wt.

[0070] The complex of polyunsaturated fatty acids in the claimed cosmetic composition includes linoleic and linolenic acids, and may also include arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid. The specified complex of polyunsaturated fatty acids has an anti-inflammatory and wound healing effect, reduces the manifestation of allergic reactions, protects the skin from UV exposure, and also helps to maintain skin moisture, thereby increasing its firmness, elasticity and reducing wrinkles. The content of the specified complex of polyunsaturated fatty acids in the claimed cosmetic composition does not exceed 2-5% wt.

[0071] Magnesium helps to maintain skin hydration. Magnesium is non-toxic, non-carcinogenic and non-comedogenic. In one embodiment, the magnesium is magnesium gluconate. In another embodiment, the magnesium is 2-pyrrolidone-5-carboxylic acid magnesium salt (MgPCA). In yet another embodiment, magnesium is a combination of magnesium gluconate and MgPCA. The total content of magnesium gluconate and MgPCA in the claimed cosmetic composition is 0.5-2% wt.

[0072] Silicon improves skin firmness by influencing the strength of elastin and collagen fibers. Silicon is included in the claimed cosmetic composition in the form of silicon dioxide. The content of silicon dioxide in the claimed cosmetic composition is 0.5-2% by weight.

[0073] In one embodiment, the claimed cosmetic composition further comprises squalane as part of an enhancer system. The content of squalane in the claimed cosmetic composition ranges from 2% wt. up to 5% wt., and the amount of water is reduced by the amount% wt., which corresponds to the content of squalane.

DESCRIPTION OF DRAWINGS

[0074] FIG. 1 schematically shows the mechanism of operation of DMSO as an enhancer of transdermal transfer.

[0075] FIG. 2 schematically shows the mechanism of NaPCa as an enhancer of transdermal transfer.

[0076] FIG. 3 illustrates the change in human skin permeability under the influence of the claimed enhancer system.

DETAILED DESCRIPTION OF THE INVENTION

[0077] In the following detailed description of the implementation of the invention, numerous implementation details are provided to provide a clear understanding of the present invention. However, it will be obvious to a person skilled in the art how the present invention can be used with or without these implementation details. In other instances, well-known techniques, procedures, and components are not described in detail so as not to obscure the details of the present invention.

[0078] In addition, it is clear from the foregoing disclosure that the invention is not limited to the foregoing implementation. Numerous possible modifications, changes, variations and substitutions, while retaining the spirit and form of the present invention, will be apparent to those skilled in the art.

[0079] The claimed system of transdermal transfer enhancers is an alternative to invasive administration of cosmetic preparations and allows delivery of biologically active substances to the dermis applied to the skin together with the claimed enhancer system or immediately after application to the skin of the claimed enhancer system. The claimed system of transdermal transfer enhancers contains DMSO, NaPCA and water in the following ratio in wt. %:

Dimethyl sulfoxide (DMSO) - 0.01-0.05,

Sodium pyrrolidone carboxylate (NaPCа) - 1-2.5,

Water - 97.45-98.99.

[0080] FIG. 1 schematically shows the mechanism of action of DMSO as an enhancer of transdermal transfer. DMSO enhances the penetration of both hydrophilic 1 BAS and lipophilic 2 BAS through the membranes of corneocytes 3 of the stratum corneum of the epidermis 4. DMSO is a good solvent and is miscible with water and organic solvents, which makes it possible to combine the claimed system of enhancers with a large number of various BAS in the composition of various cosmetics ... In addition, DMSO has anti-inflammatory, antiseptic, analgesic and fibrinolytic effects.

[0081] As a rule, the content of DMSO in the composition of cosmetics is 10% or more. However, this results in a number of side effects, which are described above. The most common side effect is dry skin, which is found in approximately 11% of the subjects [15]. One of the reasons for dry skin when using DMSO is the interaction of DMSO with keratin of corneocytes, which leads to disruption of the filamentous structure of keratin and a decrease in its ability to retain water. As noted above, dry skin weakens its protective function and can lead to a number of inflammatory conditions. It would seem that the obvious solution to this problem is to reduce the concentration of DMSO in the composition of the cosmetic product. However, at a low content, DMSO will not effectively perform its enhancer function.

[0082] In the claimed system of enhancers, a decrease in the concentration of DMSO, eliminating almost all the described side effects, is supplemented by the introduction of NaPCA as a second enhancer of transdermal transfer. In the claimed system of enhancers, NaPCA is an enhancer of the intercellular pathway of BAS penetration. FIG. 2 schematically shows the mechanism of action of NaPCA. By modifying the intercellular lipid matrix 5 of the stratum corneum of the epidermis 4 NaPCA enhances the penetration of hydrophilic 1 BAS. The combination of DMSO with NaPCA makes it possible to expand the range of biologically active substances that penetrate the epidermal barrier and, in general, enhance the conductive functions of the claimed enhancer system.

[0083] In addition to enhancing transdermal transport, NaPCA helps to increase skin hydration. NaPCA is a hygroscopic substance, its water retention capacity is more than twice that of glycerin. The increase in the NaPCA content partly compensates for the loss of water in conditions of disturbed hygroscopicity of keratin filaments. In addition, NaPCA, being a component of NMF, provides moisture to the stratum corneum of the epidermis due to the normalization of the moisturizing factor.

[0084] NaPCA in the claimed system of transdermal transfer enhancers also serves as a preservative due to its antimicrobial activity.

[0085] At a concentration not exceeding 2.5%, NaPCA is non-irritating, non-phototoxic, non-sensitizing, non-comedogen, and non-mutogen [29].

[0086] In one embodiment, the claimed system of enhancers further comprises 2-5% wt. squalane. In this case, the amount of water in the claimed enhancer system is reduced by that amount by weight. %, which corresponds to the content of squalane. Squalane has a dual function: a moisturizing agent and an enhancer of transdermal transfer. Since squalane is a form of squalene, a natural component of sebum, squalane molecules have a high affinity for the skin, mix easily with sebum, and prevent moisture from evaporating from the skin's surface. Squalane is able to penetrate further the skin surface and integrate into the intercellular lipid matrix of the stratum corneum of the epidermis. Thanks to this, squalane restores the damaged stratum corneum of the epidermis and reduces skin flaking in pathological conditions. Squalane molecules are hygroscopic, due to which they are able to retain water molecules in the skin [30].

[0087] Squalane also has transdermal transfer enhancer properties and provides an intercellular pathway for entry. Like NaPCA, it is able to change the permeability of the lipid matrix by carrying with it some BAS, such as vitamins, micro- and macroelements [3].

[0088] In addition, squalane has antioxidant properties, stimulates cell growth and skin regeneration, has antibacterial and antifungal effects [3,30]. Due to its properties, squalane increases skin elasticity, accelerates the healing of small wounds and cracks, helps smooth fine wrinkles, stimulates antioxidant processes, preventing skin aging. At the same time, squalane is non-comedogenic, practically odorless and colorless, and can be stored for more than two years without losing its functionality.

[0089] The claimed system of enhancers can be included in a cosmetic composition containing biologically active substances. In this case, the cosmetic composition has the following composition in wt. %:

Dimethyl sulfoxide (DMSO) - 0.01-0.05,

Sodium pyrrolidone carboxylate (NaPCа) - 1-2.5,

BAV - 4-10,

Water - 87.45-94.99.

[0090] In this case, the BAS in the claimed cosmetic composition include at least one peptide, and may also include a wide range of substances, for example, free amino acids and their derivatives, vitamins, polyunsaturated fatty acids, magnesium and silicon.

[0091] The claimed cosmetic composition may include either a single peptide or a combination of peptides including, but not limited to, Ala-Glu-Asp-Gly peptide, Lys-Asp-Glu peptide, Lys-Glu peptide, Arg-Lys- peptide Asp-Val-Tyr, Glu-Trp peptide, citrulline tripeptide-10, carnosine, palmitoyl pentapeptide-3, Cu-GHK peptide, palmitoyl tripeptide-38, vascular endothelial peptide (VEGF), epidermal growth factor (EGF). Moreover, the total content of peptides in the claimed cosmetic composition does not exceed 0.01-1% wt.

[0092] The peptides in the claimed cosmetic composition have a wide range of biological activities. Peptides stimulate proliferation, differentiation and activity of dermal fibroblasts, increase the expression level and quality of collagen and elastin fibrils. At the same time, peptides prevent the modification and destruction of skin proteins caused by age-related changes. In addition, peptides increase the level of skin moisture, help restore the microcircular bed, and prevent the formation of pathological scars. Thanks to this, the peptides function as geroprotectors - they help to smooth wrinkles, reduce age spots, and increase the elasticity and smoothness of the skin. In addition, peptides have an anti-inflammatory, wound healing, antiseptic effect, thereby improving the condition of the skin in case of dermatitis, erythema, infectious and inflammatory diseases. Peptides protect skin cells from the negative effects of UV, heavy metals, oxidative stress, radiation and toxic chemical factors, protect cells from chromosomal abnormalities and prevent the formation of tumors.

[0093] The Ala-Glu-Asp-Gly peptide has a wide range of biological activities. In a culture of human dermis fibroblasts, it was shown that the Ala-Glu-Asp-Gly peptide increases telomerase activity, increasing the proliferation potential of cells. This allows you to extend the period of active functioning of fibroblasts, in other words, to prolong the "cellular youth". Fibroblasts of the dermis are the main collagen-forming cells responsible for the state of the collagen framework, and, accordingly, for the elasticity and tightness of the skin. Also, the peptide Ala-Glu-Asp-Gly improves the rheological properties of blood, reduces intervascular thrombus formation. In addition, the Ala-Glu-Asp-Gly peptide is able to suppress the expression of oncogenes, thereby preventing the spontaneous appearance of tumors.

[0094] The Lys-Asp-Glu peptide has pronounced vasoprotective properties. In addition, the Lys-Asp-Glu peptide inhibits the synthesis of matrix metalloproteinases in skin fibroblasts. Due to these properties, the Lys-Asp-Glu peptide slows down the aging process of the skin and restores the capillary system, which helps nourish and moisturize the skin.

[0095] The Lys-Glu peptide regulates the rate of proliferation and differentiation of fibroblasts, stimulates their functional activity, including the synthesis of collagen and elastin. Lys-Glu peptide stimulates the skin regeneration process, thereby accelerating wound healing. Also, the Lys-Glu peptide has an immunomodulatory and oncostatic effect. In addition, the Lys-Glu peptide exhibits pronounced geroprotective properties [31–33].

[0096] Peptide Arg-Lys-Asp-Val-Tyr improves the regeneration of epidermal structures, stimulates the proliferation of epidermal cells, fibroblasts, proliferation and migration of endothelial cells (including capillary), stimulates the formation of new capillary channels, which improves blood microcirculation, nutrition and moisturizing the skin. The Arg-Lys-Asp-Val-Tyr peptide improves cutaneous immunity, thereby accelerating recovery in skin diseases such as atopic dermatitis [34, 35].

[0097] The Glu-Trp peptide has antioxidant properties, stimulating the synthesis and increasing the activity of antioxidant defense enzymes. Also, the Glu-Trp peptide stimulates the proliferation and differentiation of T-lymphocytes, regulates the production of IgA, IgG, IgM and IgE, the expression of genes of the defense systems of cells and mitochondria, and increases the activity of the nonspecific defense system. The Glu-Trp peptide has an anti-mutagenic effect, reducing the level of chromosomal aberrations when exposed to radiation and toxic-chemical factors. In addition, the Glu-Trp peptide normalizes the content of pro-inflammatory (TNF-α, IL-1β, IL-8) and anti-inflammatory (TGFβ1, IL-10) cytokines, and also inhibits the production of histamine and serotonin during inflammation. Due to this, the Glu-Trp peptide relieves inflammation and has an immunomodulatory effect, which helps to improve the condition in infectious and inflammatory skin diseases [36, 37].

[0098] Tripeptide-10 citrulline is a synthetic analogue of the skin protein decorin, the function of which is to control the formation of high-quality skin collagen. Tripeptide-10 citrulline is used for the specific targeted organization of collagen fibers. In a blind, placebo-controlled, parallel cohort study with 43 healthy volunteers, tripeptide-10 citrulline was shown to increase the uniformity of the collagen fibril diameter and increase skin elasticity due to better adhesion of collagen fibrils [38]. In addition, citrulline tripeptide-10 is able to prevent the formation of scar tissue. The formation of pathological scars and fibrous adhesions occurs mainly under the influence of the growth factor TGF-β [39]. Tripeptide-10 citrulline inhibits TGF-β activity, thereby preventing the formation of excess scar tissue.

[0099] Carnosine is a peptide with pronounced antioxidant properties [40, 41]. Carnosine is capable of chelating Cu ²⁺ and Fe ^{2+ ions} , which in free form catalyze the conversion of hydrogen peroxide into the * OH radical [42]. Also, carnosine is involved in the breakdown of reactive oxygen species (ROS) and protects the body from α-β-unsaturated aldehydes, formed from superoxidized fatty acids of cell membranes. Due to its antioxidant properties, carnosine prevents the peroxide modification of skin proteins and prevents glycation of skin proteins caused by age-related changes [43, 44]. In addition, carnosine demonstrates wound healing properties, helps smooth mimic wrinkles and prevents the development of some dermatological disorders, for example, erythema [45, 46].

[00100] Palmitoyl pentapeptide-3 enhances the expression of type I collagen and hyaluronic acid synthase-1, and also increases the secretion of procollagen and type I collagen by human fibroblasts [47]. Thanks to these properties, palmitoyl pentapeptide-3 helps to smooth wrinkles. In a double-blind, placebo-controlled, randomized trial, palmitoyl pentapeptide-3 was shown to reduce wrinkles, improve skin texture, and reduce age spots and dark circles under the eyes [48].

[00101] The Cu-GHK peptide stimulates the synthesis of collagen elastin, proteoglycan and provides anti-inflammatory and antioxidant responses. Thanks to these properties, the Cu-GHK peptide accelerates the healing of wounds, including complicated wounds such as burns, wounds with impaired blood supply and wounds that are difficult to bandage. In addition, the Cu-GHK peptide stimulates the synthesis of glycosaminoglycans - moisture-retaining molecules of the dermis, contributing to skin hydration [44, 49, 50].

[00102] Palmitoyl tripeptide-38 is a signal peptide that triggers the synthesis of such essential components of the extracellular matrix as collagen types I, III and IV, fibronectin, hyaluronic acid, laminin-5 and heat shock protein 47. Due to its signaling activity, palmitoyl tripeptide -38 promotes smoothing of wrinkles, wound healing, protection from heavy metals and oxidative stress, increases skin moisture, density and elasticity [44, 51].

[00103] Vasculoendothelial growth factor (VEGF) induces proliferation of vascular endothelial cells, enhances the formation of new blood vessels of the microcapillary bed. This improves the nutrition and hydration of the skin. In addition, VEGF affects collagen synthesis and promotes wound healing [50, 52, 53].

[00104] Epidermal growth factor (EGF) controls and stimulates the proliferation of epidermal cells, including fibroblasts. As a result, EGF promotes wound and ulcer healing when applied topically. In addition, EGF helps reduce wrinkles, age spots, skin redness, and improves skin smoothness and firmness [50, 54].

[00105] The claimed cosmetic composition as BAS may contain free amino acids and their derivatives. In this case, amino acids and their derivatives can be represented by one or more options selected from the range: glycine, alanine, valine, isoleucine, leucine, methionine, tyrosine, serine, hydroxyproline, proline, threonine, cysteine, aspartic acid, asparagine, glutamic acid, glutamine, histidine, lysine, arginine, phenylalanine, tryptophan, ectoine. Moreover, the total content of amino acids and their derivatives in the claimed cosmetic composition does not exceed 5% wt.

[00106] One of the main functions of free amino acids in the composition of the claimed cosmetic composition is moisturizing. Amino acids perform this function largely due to the fact that they are part of the NMF. In addition, free amino acids stimulate the synthesis of ceramides, and are also building blocks for collagen and elastin. Due to their properties, amino acids are involved in maintaining skin moisture, firmness and elasticity.

[00107] The vitamins in the claimed cosmetic composition prevent skin aging by reducing age-related breakdown of collagen and promoting the synthesis of collagen and elastin. In addition, vitamins help maintain skin hydration and activate enzymatic antioxidant defenses, thereby protecting the skin from sunburn. Vitamins regulate the work of the sebaceous glands, have an anti-inflammatory effect, promote the healing of wounds in minor injuries, thereby improving the condition of the skin in diseases such as acne, dermatitis, psoriasis, eczema.

[00108] The vitamins can be one or more of the following: vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, vitamin E, vitamin C and vitamin K. In this case, the total content of vitamins in the claimed cosmetic composition is 2-5% wt.

[00109] Vitamin A induces the production of type I and II procollagen and reduces age-related breakdown of collagen through inhibition of matrix metalloproteinases. The result is a thickening of the dermis and a reduction in wrinkles. In addition, vitamin A has anti-inflammatory properties, helps to restore the skin after damage, and reduce age spots [55–57].

[00110] B vitamins include vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12. B vitamins are involved in the work of the nervous system and regulate the sensitivity of the skin. In addition, B vitamins regulate the sebaceous glands and help maintain skin hydration and elasticity. Being cofactors of many enzymes, B vitamins promote the synthesis of collagen and elastin, and also exhibit antioxidant properties. B vitamins strengthen local immunity, have an anti-inflammatory effect and promote skin cell renewal. Due to their properties, B vitamins reduce wrinkles, redness, age spots, promote wound healing for minor injuries. In addition, B vitamins have a positive effect in the treatment of skin diseases such as acne, dermatitis, psoriasis, eczema [55, 56, 58] ⁠.

[00111] Vitamin E exhibits antioxidant properties. It is a versatile protector of cell membranes from oxidative damage by preventing oxygen from contacting unsaturated lipid membranes. Vitamin E also promotes the inclusion of selenium in the active center of glutathione peroxidase, thereby activating the enzymatic antioxidant defense. Vitamin E stabilizes the mitochondrial membrane, which protects mitochondria from damage. In addition, vitamin E promotes collagen production by fibroblasts. Thanks to its properties, vitamin E protects the skin from sunburn, reduces skin hyperpigmentation, edema and erythema caused by UV radiation. Also, the use of vitamin E reduces wrinkles and improves oxygen supply to skin cells [55, 59, 60].

[00112] Vitamin C has antioxidant properties, stimulates collagen synthesis, promotes the regeneration of damaged skin, enhances the protective capabilities of the epidermis against the harmful effects of UV rays. Due to its properties, vitamin C increases skin elasticity, reduces the number and depth of wrinkles, skin pigmentation.

[00113] Vitamin K has a positive effect on skin physiology, suppresses skin pigmentation, reduces redness and helps eliminate bruises [61, 62].

[00114] The complex of polyunsaturated fatty acids in the claimed cosmetic composition includes linoleic and linolenic acids, and may also include arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid. The specified complex of polyunsaturated fatty acids is appropriately called vitamin F. In the composition of the claimed cosmetic composition, the content of vitamin F is 2-5%.

[00115] Polyunsaturated fatty acids, which are part of vitamin F, are an energy substrate in the process of intracellular respiration and are part of the phospholipids of animal cell membranes. With their lack in the skin, there is a disruption in the functioning of cell membranes and fat metabolism, which leads to the development of such pathological processes as dermatitis, eczema, seborrhea, acne, cracks. Linoleic, linolenic and arachidonic acids have anti-inflammatory effects, protect the skin from UV exposure, and promote healing of wounds and ulcers.

[00116] External application of vitamin F contributes to the saturation of the skin with fatty acids, normalizes cell metabolism, promotes wound healing, reduces inflammation and the manifestation of allergic reactions. In addition, vitamin F helps to maintain skin hydration, thereby increasing its firmness, elasticity and reducing wrinkles.

[00117] Vitamin F is sensitive to high temperatures, light and contact with air, which leads to the formation of toxic oxides and free radicals, therefore it is important to use it simultaneously with antioxidants such as vitamin E or selenium.

[00118] Magnesium in the claimed cosmetic composition helps to moisturize the skin. Magnesium is an essential component of NMF and also affects the quantity and quality of fatty acids in the stratum corneum and on the surface of the epidermis. In addition, magnesium promotes collagen synthesis by skin fibroblasts [5, 63] ⁠. However, it is difficult to increase the magnesium content of the skin. With oral administration of magnesium preparations, it is largely excreted in the urine, and the remaining magnesium in the body accumulates mainly in the bones [5, 64] ⁠. In one embodiment of the cosmetic composition of the present invention, the magnesium is magnesium gluconate. In another embodiment of the cosmetic composition, the magnesium is 2-pyrrolidone-5-carboxylic acid magnesium salt (MgPCA). In yet another embodiment of the cosmetic composition, the magnesium is a combination of magnesium gluconate and MgPCA. In any specified embodiment, the total content of magnesium gluconate and MgPCA in the cosmetic composition is 0.5-2% by weight.

[00119] Silicon affects proteoglycans, complex proteins with a high degree of glycosylation. Proteoglycans are intertwined with collagen and elastin fibers, one of the main proteins of connective tissue - proteoglycan. As a result, elastin and collagen fibers are reliably held together, giving the skin elasticity. In the claimed cosmetic composition, silicon is included in the form of silicon dioxide in an amount of 0.5-2% wt.

[00120] A significant portion of these BAS have a very low affinity for the skin. The efficiency of transdermal delivery of most of these biologically active substances without additional enhancers of transdermal transfer remains a matter of scientific discussion.

EXAMPLES OF THE INVENTION

Example 1. Study of the ability of the claimed cosmetic composition containing the claimed enhancer system to change the skin permeability

[00121] In the experiment described below, a cosmetic composition was investigated comprising an enhancer system containing 0.01 wt. DMSO and 2% wt. NaPCA. As a test object, a three-dimensional model of human skin was used, consisting of a reconstructed epidermis with a functional stratum corneum "VitroSkin" (developed by OOO NPO VitroLab, corresponds to paragraph 4.2 of GOST 32634-2014). Setting up an experiment with the study of the dynamics of penetration of a control substance through the human epidermis in an in vitro test allows one to give an integral assessment of the change in skin permeability under the influence of the tested product [65]. Fluorescein (CAS Number: 2321-07-5), a low-molecular-weight organic compound capable of diffusion across the skin barrier, was used as a control substance, and a fluorescence method was used as the physical principle of detection. 0.9% NaCl was used as a negative control. The test substance (cosmetic composition or negative control substance) was applied topically to a VitroSkin skin model. The ability of the test substance to change the skin permeability was determined by the amount of fluorescein penetrated from the apical part of the transwell to the basolateral part. The amount of fluorescein penetrated was determined from the fluorescence intensity of the solution in the basolateral region, determined using a VarioSkan fluorometer (ThermoFisher). First, the dynamics of fluorescein penetration through the thickness of the skin model was investigated. For this, 100 μl of fluorescein was applied to a skin model sample (in the apical part of the transwell). Immediately after application (0 minutes), after 30 and 60 minutes, 50 μl were taken from the basolateral section. The withdrawn volume was replenished with the appropriate amount of 0.9% NaCl. For each exposure time, at least two skin model samples were used. Then, the dynamics of fluorescein penetration through the thickness of the skin model was investigated after exposure of its surface to the test cosmetic composition or negative control substance. The test substance was applied uniformly for 3 minutes on the surface of a skin model sample with a full coverage of at least 25 μl / cm ² . At the end of the exposure period, the test substance was thoroughly washed off the surface of the skin model with 0.9% NaCl solution. Then 100 μl of fluorescein was applied to a skin model sample (in the apical part of the transwell). Immediately after application (0 minutes), after 30 and 60 minutes, 50 μL of solution was taken from the basolateral section. The withdrawn volume was replenished with the appropriate amount of 0.9% NaCl. For each exposure time, at least two skin model samples were used.

[00122] The measurement results are shown in Table 1 and also illustrated in FIG. 3.

Table 1. Dynamics of fluorescein penetration through the thickness of the skin model sample without exposure and after exposure to its surface of a cosmetic composition containing the claimed enhancer system, or a negative control substance

No. Processing option Exposure time, min Fluorescence intensity, RFU 1 Without processing 0 0 2 Without processing 0 0 3 Without processing 0 0 4 Without processing thirty 4.22 five Without processing thirty 9.69 6 Without processing thirty 6.85 7 Without processing 60 21.99 eight Without processing 60 23.45 nine Without processing 60 20.35 ten Cosmetic composition 0 0 eleven Cosmetic composition 0 0 12 Cosmetic composition 0 0 13 Cosmetic composition thirty 12.76 fourteen Cosmetic composition thirty 14.34 fifteen Cosmetic composition thirty 11.01 sixteen Cosmetic composition 60 16.55 17 Cosmetic composition 60 19.86 18 Cosmetic composition 60 15.52 nineteen Negative control 0 0 20 Negative control 0 0 21 Negative control 0 0 22 Negative control thirty 3.57 23 Negative control thirty 4.22 24 Negative control thirty 3.05 25 Negative control 60 9.98 26 Negative control 60 7.23 27 Negative control 60 6.86

[00123] FIG. 3 are graphs showing the kinetics of fluorescein penetration through the thickness of the skin model without any effect on the skin model sample and after exposure of the skin model sample to the test and control substances. Graph 6 depicts the dynamics of fluorescein penetration through the thickness of an untreated skin model sample. Graph 7 was obtained by processing a skin model sample of the tested cosmetic composition, graph 8 - by treatment with a negative control substance. The dots on the graphs show the arithmetic mean for each exposure time. It can be seen that 30 min after exposure, the penetration of fluorescein through the thickness of the skin model is noticeably stronger after exposure to the tested cosmetic composition, compared with exposure to a negative control substance or without any exposure. As shown by the statistical analysis using the Student's test, the differences observed between the effect of the tested cosmetic composition and the negative control substance were significant both for an exposure time of 30 min ( p <0.01) and for an exposure time of 60 min ( p <0.01 ). Also, at 30 minutes after exposure, the penetration of fluorescein through the thickness of the skin model sample treated with the test cosmetic composition was significantly different from the penetration through the untreated sample ( p <0.05). At the same time, 60 minutes after exposure, the penetration of fluorescein through the thickness of the skin model did not differ for the sample treated with the cosmetic composition and the untreated sample. Based on the results of the experiment, it was concluded that a cosmetic composition containing the claimed system of enhancers significantly increases skin permeability 30 minutes after exposure. At the same time, after 60 minutes, skin permeability returns to normal, which is critical for maintaining healthy skin.

Example 2. Study of the ability of the claimed cosmetic composition containing the claimed enhancer system to cause skin irritation

[00124] In the experiment described below, a cosmetic composition was investigated comprising an enhancer system containing 0.01% wt. DMSO and 2% wt. NaPCA. A three-dimensional model of human skin, consisting of a reconstructed epidermis with a functional stratum corneum "VitroSkin"("VitroLab"), was used as a test object. The research protocol was formed in accordance with GOST 32634-2014. The study according to this protocol allows to reliably identify the presence or absence of the ability of the tested substances to cause irritation / damage to the skin [66–68]. The test substance was applied to the surface of the skin model. The corrosive properties of a substance were determined by its ability to cause a decrease in the viability below the threshold level for certain periods of exposure. Cell viability was determined using the MTT test [69]. The principle of the skin model study is based on the assumption that corrosive chemicals can penetrate into the stratum corneum by diffusion or erosion and have a cytotoxic effect on the underlying cell layers. To confirm the proper operation of the experimental models, an aqueous solution of 0.9% by weight was used as a negative control substance. NaCl, an aqueous solution of 8N KOH was used as a positive control. The test substance was applied to the surface of the skin model sample so that it covered at least 25 μl / cm ^{2 of the} surface of the skin model sample. The exposure time was 3 minutes and 60 minutes. At the end of the exposure period, the test substance was thoroughly washed off the surface of the skin model sample with a 0.9% wt. NaCl. To assess cell viability, a validated quantitative method was used that is compatible with the 3D skin model and provides accurate and reproducible results (Fentem et al., 1998). This method is based on the ability of mitochondrial enzymes to reduce the MTT dye [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide] to insoluble formazan. A skin model sample was placed in a 1 mg / ml MTT solution and kept at a temperature of 37 ° C for 3 hours. Then the precipitated formazan was extracted with isopropanol and the concentration of the formazan was determined by measuring the optical density at a wavelength of 595 nm. The optical density measurement results for all samples are presented in Table 2.

[00125] Table 2. Optical density of the formazone extract

No. Sample type Optical density value 1 Negative control 0.675 2 Negative control 0.620 3 Negative control 0.719 4 Positive control 0.090 five Positive control 0.067 6 Positive control 0.105 7 Cosmetic composition, exposure 3 min 0.611 eight Cosmetic composition, exposure 3 min 0.779 nine Cosmetic composition, exposure 3 min 0.5 ten Cosmetic composition, exposure 60 min 0.489 eleven Cosmetic composition, exposure 60 min 0.470 12 Cosmetic composition, exposure 60 min 0.568

The absorbance values for each substance and exposure time were averaged, and the average absorbance values were used to calculate the percent viability versus a negative control, which was arbitrarily set at 100%. Data on cell viability after exposure to the test substances are presented in Table 3.

[00126] Table 3 . Cell viability after exposure to the claimed cosmetic composition containing the claimed enhancer system

No. Sample type Average viability,% Average error,% 1 Negative control 100 5.16 2 Positive control 13.01 1.64 3 Cosmetic composition,
exposure 3 min 93.84 12.08 4 Cosmetic composition,
exposure 60 min 75.82 4.47

[00127] In the method used, the pot life threshold after 3 minutes exposure is 50%, the pot life threshold after 60 minutes exposure is 15%. Table 4 shows that the cell viability after 3 minutes of exposure to the tested cosmetic composition was 93.84%, and after 60 minutes of exposure - 75.82%, which is significantly higher than the threshold values. Based on the experimental data obtained, it was concluded that the claimed system of enhancers does not cause irritation / damage to human skin.

[00128] In the present application materials, the preferred disclosure of the implementation of the claimed technical solution is presented, which should not be used as limiting other, particular embodiments of its implementation, which do not go beyond the scope of the claimed scope of legal protection and are obvious to specialists in the relevant field of technology.

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Claims (16)

1. The system of enhancers of transdermal transfer, characterized in that it contains dimethyl sulfoxide and sodium pyrrolidone carboxylate, while the ratio of components in wt. %: dimethyl sulfoxide 0.01-0.05 sodium pyrrolidone carboxylate (NaPCA) 1-2.5 water - the rest 2. The system of transdermal transfer enhancers according to claim 1, additionally containing squalane, wherein the ratio of components in wt. %: dimethyl sulfoxide 0.01-0.05 sodium pyrrolidone carboxylate (NaPCA) 1-2.5 squalane 2-5 water - the rest 3. Cosmetic composition containing a system of transdermal transfer enhancers, including dimethyl sulfoxide and sodium pyrrolidone carboxylate, as well as biologically active substances, including at least one peptide, while the ratio of components in wt. %: dimethyl sulfoxide 0.01-0.05 sodium pyrrolidone carboxylate 1-2.5 biologically active substances 4-10 in the water rest 4. Cosmetic composition according to claim 3, where the peptide is selected from the series: epitalon, peptide KDE, peptide KE, peptide RKDVY, peptide EW, tripeptide-10 citrulline, carnosine, palmitoyl pentapeptide-3, peptide Cu-GHK, palmitoyl tripeptide-38 , vascular endothelial growth factor, epidermal growth factor. 5. Cosmetic composition according to claim 3, characterized in that the biologically active substances comprise at least one vitamin. 6. Cosmetic composition according to claim 5, where the vitamin is selected from the series: vitamin A, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, vitamin E, vitamin C, vitamin K ... 7. A cosmetic composition according to claim 3, characterized in that the biologically active substances comprise at least one amino acid or an amino acid derivative. 8. Cosmetic composition according to claim 7, wherein the amino acid or amino acid derivative is selected from the range: glycine, alanine, valine, isoleucine, leucine, methionine, tyrosine, serine, hydroxyproline, proline, threonine, cysteine, aspartic acid, asparagine, glutamic acid, glutamine, histidine, lysine, arginine, phenylalanine, tryptophan, ectoine. 9. Cosmetic composition according to claim 3, characterized in that the biologically active substances include a complex of polyunsaturated fatty acids containing linoleic acid and linolenic acid. 10. Cosmetic composition according to claim 3, characterized in that the biologically active substances include magnesium. 11. Cosmetic composition according to claim 3, characterized in that the biologically active substances include silicon. 12. Cosmetic composition according to p. 3, characterized in that the system of transdermal transfer enhancers additionally includes squalane, while the ratio of components in wt. %: dimethyl sulfoxide 0.01-0.05 sodium pyrrolidone carboxylate 1-2.5 squalane 2-5 biologically active substances 4-10 purified water - the rest

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