RU2792212C1 - Composition for skin protection from exposure to harmful substances, light and stress - Google Patents

Abstract

FIELD: protection of the skin from environmental exposure.

SUBSTANCE: group of inventions relates to a composition for the protection of the skin from environmental exposure. Cosmetic composition for the protection of the skin from exposure to harmful substances, light, or combinations thereof, contains peptides and amino acids, where peptides include nicotinoyl tripeptide, pentapeptide, nicotinoyl hexapeptide, sh-octapeptide, tetracarboxymethyl hexanoyl dipeptide, hexacarboxymethyl dipeptide and biotinoyl hexapeptide-2 amide, and amino acids include arginine, asparagine, aspartic acid, glycine, histidine, hydroxyproline, isoleucine, leucine, methionine, phenylalanine, proline, serine and valine. Also, a pharmaceutical composition for the prevention or treatment of diseases caused by exposure to harmful substances, light, or a combination thereof, and a filler composition for protecting the skin from exposure to harmful substances, light, or a combination thereof are described.

EFFECT: group of inventions provides skin protection by inhibiting the abnormal cell response to external stress stimuli, such as harmful substances and light, without any cytotoxicity.

16 cl, 10 dwg, 1 tbl, 6 ex

Description

Technical field

The present invention relates to compositions for protecting the skin from the effects of stress caused by environmental conditions such as harmful substances and light.

State of the art

As a rule, human skin is divided into epidermis, dermis and hypodermis; The epidermis is subdivided into the stratum corneum, the stratum stratum lucidum (cycloid layer), the granular layer, the spiny layer, and the basal layer. The stratum corneum, the outer layer of the skin, is made up of dead keratinocytes. It is essentially composed of keratinocytes, which in turn are composed of protein, with the space between the keratinocytes formed by lipids such as ceramide, cholesterol, and free fatty acids. These lipid layers prevent the evaporation of moisture from the body and have both a healing function, protecting the body from external irritants, and an aesthetic function, giving the skin a shiny and smooth appearance.

The skin is directly exposed to the external environment and, under the influence of harmful substances existing in the external environment, becomes prone to irritation and inflammatory reactions such as erythema, swelling and itching. Contact with harmful substances causes stimulation and an inflammatory reaction due to toxins, whereby substances can cause skin pigmentation, reduce skin elasticity and contribute to damage to the skin barrier.

It is widely known that UV rays can accelerate skin damage and aging. In particular, when the skin is overexposed to ultraviolet rays, free radicals accumulate to form active oxygen. This causes oxidative stress and destroys the antioxidant system, causing damage to the skin.

In addition, the blue light emitted by displays of smart devices such as TV and computer, as well as LED lighting products, is blue light with a wavelength of 380 to 500 nanometers (nm), which has the shortest and strongest wavelength among visible light. . There are studies that skin exposure to blue light causes pigmentation (Luc Duteil et al. Pigment Cell Melanoma Res 2014; 27: 822-6).

Physical stimulation from the environment can cause cancer due to DNA damage in cells and skin, or can cause damage to the central and peripheral nervous system, destruction of the immune system, reproductive system disorders, inflammatory diseases, chronic allergies, and the like.

Thus, there is a need to develop a composition that is safe and has no side effects, while protecting the skin from external influences such as harmful substances and ultraviolet rays.

Detailed description of the invention

Technical task

The object of the present invention is to provide a composition for protecting the skin from stress caused by external influences such as harmful substances and light.

The solution of the problem

To solve the above problems, the present invention proposes a composition for protecting the skin from exposure to harmful substances and light, while this composition contains:

peptides and amino acids,

wherein the peptides include nicotinoyl tripeptide, pentapeptide, nicotinoyl hexapeptide, sh-octapeptide, tetracarboxymethyl hexanoyl dipeptide, hexacarboxymethyl dipeptide and biotinoyl hexapeptide-2 amide, and

amino acids include arginine, asparagine, aspartic acid, glycine, histidine, hydroxyproline, isoleucine, leucine, methionine, phenylalanine, proline, serine, and valine.

According to one implementation variant, the composition according to the present invention may further contain hyaluronic acid, while the molecular weight of hyaluronic acid may be from 500 kDa to 2 MDa.

In one embodiment, the harmful substances may include one or more substances selected from the group consisting of yellow dust, fine dust, cigarette smoke, vehicle exhaust, heavy metals, and air pollutants.

According to another implementation variant of the specified composition can inhibit the death of skin cells caused by stress from exposure to harmful substances and light.

In addition, said composition may have antioxidant activity.

In addition, said composition can inhibit the formation of cyclobutane pyrimidine dimers (CPDs).

In addition, said composition can adsorb or remove harmful substances.

In addition, said composition can be used to smooth wrinkles or improve elasticity.

According to another implementation variant, the peptides may contain, based on 100 parts by weight of sh-octapeptide,

from 1 to 50 mass parts of nicotinoyl tripeptide;

from 10 to 150 mass parts of the pentapeptide;

from 0.05 to 10 mass parts of nicotinoyl hexapeptide;

from 5 to 70 mass parts of tetracarboxymethyl hexanoyl dipeptide;

from 15 to 80 mass parts of hexacarboxymethyl dipeptide; And

5 to 70 parts by weight of biotinoyl hexapeptide-2 amide, and

amino acids may contain, based on 100 mass parts of valine,

from 80 to 180 mass parts of arginine;

from 10 to 60 mass parts of asparagine;

from 20 to 80 mass parts of aspartic acid;

from 30 to 100 mass parts of glycine;

from 50 to 150 mass parts of histidine;

from 5 to 40 mass parts of hydroxyproline;

from 40 to 120 mass parts of isoleucine;

from 50 to 150 mass parts of leucine;

from 5 to 40 mass parts of methionine;

from 40 to 120 mass parts of phenylalanine;

from 10 to 50 mass parts of proline; And

from 20 to 80 mass parts of serine.

According to a variant of one implementation, the composition may additionally contain from 1 to 5% of the mass. hyaluronic acid based on the total weight of the composition.

In another aspect of the present invention, there is provided a cosmetic composition for reducing skin darkening, wrinkles, or improving elasticity caused by exposure to harmful substances and light, comprising the above-described composition.

In one embodiment, the cosmetic composition may contain non-crosslinked hyaluronic acid.

Moreover, in another aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating diseases caused by exposure to harmful substances and light, comprising the composition as described above. The disease may be selected from the group consisting of cancer, atopic dermatitis, contact dermatitis, seborrheic dermatitis, erythema, sunburn, photosensitivity, acne, dry skin, psoriasis, lethality, immunotoxicity, peripheral nerve injury, central nerve injury, endocrine gland injury, reproductive disorders, developmental disorders in infants and young children, anemia, bronchitis, emphysema, decreased lung function, asthma, arrhythmia, heart attack, angina pectoris and myocardial infarction.

In one embodiment, the pharmaceutical composition of the present invention may be presented as an injectable filler.

According to another implementation variant, the pharmaceutical composition may contain cross-linked hyaluronic acid.

Specific details of other embodiments of the present invention are included in the following detailed description of the invention.

Benefits of the Invention

The present invention protects the skin by inhibiting the abnormal response of cells to external stress stimuli such as harmful substances and light without any cytotoxicity, so that it can be effectively applied in various fields such as pharmaceuticals and cosmetics.

Brief description of graphic materials

On FIG. 1 is a diagram showing cell viability under blue light.

On FIG. 2 shows an optical micrograph of the observed changes in skin tissues under the influence of ultraviolet rays.

On FIG. 3 and 4 are diagrams and photomicrographs showing the formation of cyclobutane pyrimidine dimers (CPDs), respectively.

On FIG. 5 is a diagram showing cell viability under the influence of harmful substances.

On FIG. 6 is a diagram showing cell viability under the influence of harmful substances and ultraviolet radiation.

On FIG. 7 is a diagram showing the ability to scavenge free radicals under the influence of harmful substances and ultraviolet rays.

On FIG. 8 - Fig. 10 are photographs and diagrams showing the wrinkle-smoothing effect of injecting the composition of the present invention.

The best embodiments of the invention

Since various changes and variations may be made to the present invention, specific embodiments are illustrated in the drawings and will be described in detail in the detailed description of the invention. However, it should be understood that the present invention is not intended to be limited to specific embodiments, and includes all modifications, equivalents, and alternatives falling within the spirit and scope of the present invention. In the following description of the present invention, a detailed description of the prior art will be omitted if it is found that this may obscure the essence of the present invention.

Unless otherwise specified in the present description of the invention, the expression "between" is used as an expression that includes the corresponding numerical value. In particular, for example, the expression "from 1 to 2" implies the inclusion of all numbers between 1 and 2, as well as 1 and 2.

Hereinafter, the skin protection composition according to an embodiment of the present invention will be described in more detail.

In the context of the present invention, the term "protection of the skin" means to help prevent, improve or treat damage to the skin from the harsh conditions to which it may be exposed from the external environment. Harsh conditions include, in particular, harmful substances or light.

For example, the harmful substances may contain one or more substances selected from the group consisting of yellow dust, fine dust, cigarette smoke, vehicle exhaust, heavy metals, and air pollutants.

In addition, the term "light" in the context of the present invention means ultraviolet light, blue light, or a combination thereof, unless otherwise indicated.

Air pollutants may include substances and compounds listed in section 4 of the Clean Air Preservation Act (as amended in 2019). In particular, air pollutants may include one or more substances selected from the group consisting of cadmium and its compounds, hydrogen cyanide, lead and its compounds, polychlorinated biphenyls, chromium and its compounds, arsenic and its compounds, mercury and its compounds, propylene oxide , chlorine and its compounds, fluorine compounds, asbestos, nickel and its compounds, vinyl chloride, dioxin, phenol and its compounds, beryllium and its compounds, benzene, carbon tetrachloride, methyl disulfide, aniline, chloroform, formaldehyde, acetaldehyde, benzidine, 1, 3 -butadiene, polycyclic aromatic hydrocarbons, ethylene oxide, dichloromethane, styrene, tetrachlorethylene, 1,2-dichloroethane, ethylbenzene, trichlorethylene, acrylonitrile and hydrazine.

The present invention provides a composition for protecting the skin from stress caused by exposure to harmful substances and light, containing peptides.

In particular, the present invention includes nicotinoyl tripeptide, pentapeptide-37, nicotinoyl hexapeptide, sh-octapeptide, tetracaroxymethyl hexanoyl dipeptide, hexacarboxymethyl dipeptide and biotinoyl hexapeptide-2 amide.

Nicotinoyl hexapeptide is a product derived from the interaction of vitamin B3 and hexapeptide, which activates the skin and helps to reduce skin hypersensitivity.

Pentapeptide helps to whiten and moisturize the skin.

Nicotinoyl hexapeptide is a product obtained from the interaction of vitamin B3 and hexapeptide, which reduces the function of PAR2 (signal transduction receptor) that causes exogenous aging, and also helps to prevent oxidation and protect the skin.

Sh-octapeptide is a single chain recombinant human peptide containing 8 amino acids consisting of glycine, histidine, leucine, lysine, phenylalanine and tyrosine and prevents oxidative damage to the skin.

Tetracarboxymethyl hexanoyl dipeptide and hexacarboxymethyl dipeptide are used as a skin conditioning agent.

Biotinoyl hexapeptide-2 amide is a reaction product of hexapeptide-2 and biotin, wherein the carboxyl end (C-terminus) of the peptide is an amide. It is used as a skin conditioning agent.

According to an implementation variant, the peptides may contain, based on 100 mass parts of sh-octapeptide,

from 1 to 50 mass parts, for example, from 5 to 30 mass parts, for example, from 10 to 30 mass parts of nicotinoyl tripeptide;

from 10 to 150 mass parts, for example, from 30 to 100 mass parts, for example, from 50 to 70 mass parts of the pentapeptide;

from 0.05 to 10 mass parts, for example, from 0.1 to 5 mass parts, for example, from 0.5 to 4 mass parts of nicotinoyl hexapeptide;

from 5 to 70 mass parts, for example, from 10 to 50 mass parts, for example, from 20 to 40 mass parts of tetracarboxymethyl hexanoyl dipeptide;

from 15 to 80 mass parts, for example, from 30 to 60 mass parts, for example, from 40 to 50 mass parts of hexacarboxymethyl dipeptide; And

5 to 70 parts by weight, eg 10 to 50 parts by weight, eg 20 to 40 parts by weight of biotinoyl hexapeptide-2 amide.

According to one implementation variant, the present invention may further include amino acids, in particular, amino acids may further include one or more amino acids selected from the group consisting of arginine, asparagine, aspartic acid, glycine, histidine, hydroxyproline, isoleucine, leucine, methionine, phenylalanine , proline, serine and valine, for example.

In particular, amino acids may include, based on 100 mass parts of valine,

from 80 to 180 mass parts, for example, from 50 to 250 mass parts, for example, from 70 to 150 mass parts, for example, from 90 to 130 mass parts of arginine;

from 10 to 60 mass parts, for example, from 20 to 50 mass parts, for example, from 25 to 40 mass parts of asparagine;

from 20 to 80 mass parts, for example, from 30 to 70 mass parts, for example, from 35 to 50 mass parts of aspartic acid;

from 30 to 100 mass parts, for example, from 40 to 80 mass parts, for example, from 40 to 60 mass parts of glycine;

from 50 to 150 mass parts, for example, from 60 to 120 mass parts, for example, from 70 to 100 mass parts of histidine;

from 5 to 40 mass parts, for example, from 10 to 30 mass parts, for example, from 10 to 20 mass parts of hydroxyproline;

from 40 to 120 mass parts, for example, from 50 to 100 mass parts, for example, from 60 to 80 mass parts of isoleucine;

from 50 to 150 mass parts, for example, from 60 to 120 mass parts, for example, from 70 to 100 mass parts of leucine;

from 5 to 40 mass parts, for example, from 10 to 30 mass parts, for example, from 10 to 20 mass parts of methionine;

from 40 to 120 mass parts, for example, from 50 to 100 mass parts, for example, from 60 to 80 mass parts of phenylalanine;

from 10 to 50 mass parts, for example, from 20 to 40 mass parts, for example, from 20 to 30 mass parts of proline; And

20 to 80 parts by weight, eg 30 to 60 parts by weight, eg 35 to 50 parts by weight serine.

The composition of the present invention can protect skin cells by adsorbing or releasing harmful substances.

In addition, it can inhibit skin damage caused by ultraviolet rays. DNA damage caused by ultraviolet radiation can occur in the skin, leading to the formation of cyclobutane pyrimidine dimers (CPDs). In CPE, the double bonds in the hexagonal ring structures of adjacent pyrimidine bases are transformed by exposure to short wavelength ultraviolet light (wavelength about 254 nm) to form a new tetragonal bond that causes abnormal binding causing structural damage in DNA. The composition according to the present invention can inhibit the formation of CPE, thus inhibiting apoptosis of skin cells caused by exposure to ultraviolet rays.

In addition, it is possible to increase the antioxidant activity, prevent and alleviate damage to skin cells caused by the stress of exposure to harmful substances and light, and thereby reduce skin darkening, smooth wrinkles, or improve skin elasticity.

In one embodiment, the composition of the present invention may be formulated into any dosage form suitable for dermatological use, such as a conventional cosmetic composition, a topical pharmaceutical composition, and the like. A composition suitable for use in dermatology refers to a composition that may have a relatively non-toxic and harmless effective effect on the subject and may contain a topical preparation that can be applied to the skin, the composition does not impair the activity and properties of the active ingredient, does not causing side effects due to the composition and does not reduce the effectiveness of the active ingredient or cause serious irritation in the subject.

Moreover, the external composition for skin according to the present invention can be prepared in any dosage form accepted in the art, exemplified by solution, suspension, liquid, emulsion, paste, gel, mass, cream, lotion, powder, but not limited to soap, surfactant-containing cleanser, oil, cream powder foundation, emulsion base, wax base, spray, and hair cosmetics.

Specifically, the cosmetic composition can be provided as a skin lotion, skin emollient, skin tonic, pore reducer, lotion, body milk, moisturizing lotion, nourishing lotion, massage cream, nourishing cream, moisturizing cream, cream. for hands, base cream, essence, nourishing essence, mask, soap, hair shampoo, foot shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser, etc., and, for example, can be obtained as a cosmetic filler.

The cosmetic filler can be used in a method of application to a skin surface. It may include perfume, xanthan gum, wax, solid oil, liquid oil, surfactant, moisturizer, alcohol, and it may be included without particular limitation as long as it is a component of a commonly used composition. In addition, if the cosmetic filler composition contains hyaluronic acid, the hyaluronic acid may have a non-crosslinked structure.

In addition, in another embodiment, the present invention provides a pharmaceutical composition for preventing or treating diseases caused by harmful substances and light. In particular, the diseases may be selected from the group consisting of cancer, atopic dermatitis, contact dermatitis, seborrheic dermatitis, erythema, sunburn, photosensitivity, acne, dry skin, psoriasis, lethality, immunotoxicity, peripheral nerve injury, central nerve injury, endocrine glands, reproductive system disorders, developmental disorders in infants and young children, anemia, bronchitis, emphysema, decreased lung function, asthma, arrhythmia, heart attack, angina pectoris and myocardial infarction.

According to one implementation variant of the pharmaceutical composition according to the present invention may contain a pharmaceutically acceptable carrier. A pharmaceutically acceptable carrier includes binders, glidants (lubricants), disintegrants, excipients, lubricants, solvents, dispersing agents, stabilizers, suspending agents, coloring agents, flavoring agents, buffers, preservatives, emollients, anesthetics, isotonic agents, or their combinations. The dosage form of the pharmaceutical composition according to the present invention can be obtained in various ways by mixing with a pharmaceutically acceptable carrier, examples of which are described above. The pharmaceutical composition according to the present invention can be prepared as medical auxiliary materials, solution, suspension, ointment, tablet, capsule, etc., but the present invention is not limited to them.

Medical ancillary materials may include medical fillers, wound dressings, bone grafts, soft tissue adjuvants, dental materials, etc., which can be used to create artificial tissues, artificial organs, cell culture scaffolds, drug delivery agents, biosensors for transplantation, etc.

In one embodiment, the composition of the present invention may be formulated as an injectable filler which may contain one or more carriers selected from the group consisting of alginic acid, carboxymethyl cellulose, chitosan, dextran, collagen, gelatin, pectin, agar, amylose, cyclodextrin and elastin. In addition, if the injectable filler composition contains hyaluronic acid, the hyaluronic acid may have a cross-linked structure.

Further, for example, the biodegradable polymer scaffold may include hyaluronic acid, polyglycolic acid (PGA), polylactide (PLA), polylactide-glycolic acid copolymer (PLGA), polycaprolactone (PCL), polyamino acid, polyanhydride, polyorthoester, or a copolymer thereof.

According to another implementation variant, the composition according to the present invention contains hyaluronic acid. Hyaluronic acid may include a salt or cross-linked hyaluronic acid, such as sodium hyaluronate. In addition, for example, the molecular weight of hyaluronic acid may be from 500 KDa to 2 MDa, for example, from 1 to 2 MDa, for example, from 1 to 1.5 MDa. The content of hyaluronic acid in the present invention may be from 1 to 5% wt., for example, from 1 to 3% of the mass. based on the total weight of the composition.

In another embodiment, the biodegradable polymer scaffold may have pores formed therein, wherein the pores are formed by salts, effervescent salts, carbohydrates, hydrocarbon waxes, and the like.

Further, the injectable filler of the present invention may include a local anesthetic, an antihistamine, vitamins, and the like.

The local anesthetic may include, for example, lidocaine, etidocaine, bupivacaine, tetracaine, mepivacaine, procaine, prilocaine, ropivacaine, etc., and is not particularly limited when said agent is an injectable local anesthetic.

The antihistamine may include, for example, piprylhydrinate (Plokon), chlorphenamine, diphenylpyralin, diphenhydramine, cetirizine, etc., and is not particularly limited when said agent is an injectable antihistamine.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily carry out the present invention. However, the present invention may be embodied in various forms and is not limited to the embodiments described herein.

Examples

The skin protection composition was obtained from the composition shown in Table 1.

Table 1 Substance Composition % mg/kg Peptides Nicotinoyl tripeptide-1 0.00012 1.2 Pentapeptide-37 0.00040 4.0 Nicotinoyl hexapeptide-44 0.00001 .0.1 sh-octapeptide-4 0.00060 6.0 Tetracarboxymethyl hexanoyl dipeptide-12 0.00020 2.0 Hexacarboxymethyl dipeptide-12 0.00029 2.9 Biotinoyl hexapeptide-2 amide 0.00020 2.0 Amino acids Arginine 0.00062 6.2 Asparagine 0.00016 1.6 Aspartic acid 0.00020 2.0 Glycine 0.00028 2.8 Histidine 0.00040 4.0 Hydroxyproline 0.00008 0.8 Isoleucine 0.00036 3.6 Leucine 0.00040 4.0 Methionine 0.00008 0.8 Phenylalanine 0.00034 3.4 Proline 0.00012 1.2 Serene 0.00022 2.2 Valine 0.00050 5.0 Hyaluronic acid as sodium hyaluronate 1.50000 15000 Water up to 100

Example 1: protecting cells from exposure to blue light

To confirm the cytoprotective effect of the composition according to the present invention against exposure to blue light MTS study , cell viability analysis was performed using human fibroblasts (NHDFs).

Human fibroblast cells (Normal Human foreskin-derived Dermal Fibroblasts, NHDFs; ATCC, CRL-2522) at a concentration of 3×104 cells/well were placed in a 24-well plate and cultured in a modified solution of Dulbecco's medium (DMEM) containing 10% fetal bovine serum and 100 U/ml penicillin/streptomycin solution at 5% CO ₂ and 37°C for 24 hours. The Example formulation was prepared at a concentration of 0 to 1.0% and cultured cells were treated with the formulation for 24 hours and then irradiated with blue light (453 nm, 40 J/cm ² ) for 6 hours to measure cell viability. For measurement, after removal of the culture medium, the cells were treated with a solution of MTS (0.33 mg/ml MTS) for 1 to 4 hours, the absorbance was measured at a wavelength of 490 nm using a spectrophotometer (Epoch 2, BioTek).

The results of the quantitative determination of MTS, the protective effect of human fibroblasts under blue light, are shown in FIG. 1. As a result, in terms of cell viability under blue light irradiation, the group treated with the composition of the present invention showed 1 to 11% higher cell viability compared to the group irradiated with blue light and not treated with the composition.

Example 2: Prevention of skin damage caused by ultraviolet rays (ex vivo)

To confirm the prophylactic effect of the composition of example (product S) on skin injury, skin biopsy samples treated with the composition of example were irradiated with ultraviolet light (UV-A/UV-B) to measure the formation of skin explants (H/E Staining) and the amount of cyclobutane pyrimidine dimers. (CPDs). Ultraviolet radiation causes DNA damage, which is due to the formation of CPE.

An ex vivo model using full-thickness human skin tissues has been approved by the ethical committee with informed consent. The selected skin tissues were cultured for 24 hours under the following conditions: CO ₂ content of 5% and temperature of 37°C, after which measurements were taken.

Skin tissue explants were cultured in a 12-well plate for 24 hours, then a sample (example composition) was prepared at a concentration of 30%, and the cultured cells were treated with the sample for 24 hours and irradiated with ultraviolet rays (UV-A/UV-B) for 5 o'clock. The shape of the skin tissues (6 μm thick section) was observed using an optical microscope (DM2000, 20X objective) by staining with hematoxylin/eosin (H/E staining) (FIG. 2).

The tissue sample was then treated under the same CPE determination conditions as in the example described above, and then irradiated with ultraviolet rays (UV-A/UV-B) for a certain period of time (30 min, 1 h 30 min, 5 h).

To measure the representation of the CPP used immunohistochemical study (IHC) embedded in paraffin tissue section (Fig. 3), tissue samples were examined using an optical microscope (Fig. 4). The dose of UV radiation A was 50 J/cm2, the dose of UV radiation B was 500 mJ/cm2.

On FIG. 2, the thickness of the epidermal layer and the density of the dermal layer were evaluated as an indicator of photoaging when exposed to UV rays. As a result of skin tissue observation, it was confirmed that in the group exposed to UV irradiation and not treated with the composition of Example, the epidermal layer was thickened and the density of the dermal layer was reduced. At the same time, it was found that in the group treated with the composition of Example (Product S), the epidermal layer had a small thickness, and the density of the dermal layer was increased.

On FIG. 3, the representation of CPE showed an average decrease of 3 to 7% in the UV irradiated group treated with the example composition compared to the UV irradiated group not treated with the example composition.

In the process of observing the formation of CPP in skin tissues in Fig. 4, the formation of CPP was clearly observed in the group exposed to UV irradiation and not treated with the composition of the example. At the same time, in the group exposed to UV irradiation and treated with the composition of example, a decrease in the tendency to form CPE was observed compared to the group irradiated with UV rays and not treated with the composition of example.

Example 3: Protecting cells from exposure to air pollutants (in vivo)

To confirm the cytoprotective effect of the composition according to the present invention against air pollutants, human fibroblasts (NHDFs) were treated with urban dust as an air pollutant and the composition of the example. Cell viability was assessed by determining MTS ((3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazole).

As urban dust, we used (SRM1649b, US National Institute of Standards and Technology (NIST)) atmospheric particles sampled in Washington in the period from 1976 to 1977. Urban dust contains many compounds such as polycyclic aromatic hydrocarbons (PAHs), nitro-substituted polycyclic aromatic hydrocarbons, polychlorinated biphenyl (PCB) congeners, chlorinated pesticides, decabromodiphenyl ether (DecaBDE), dibenzo-p-dioxin and dibenzofuran congeners. The particle size was approximately 10 μm.

Human fibroblast cells (normal human foreskin-derived dermal fibroblasts, NHDFs; ATCC, CRL-2522) were placed in a 24-well plate at a concentration of 3×10 ⁴ cells/well and cultured in DMEM containing 10% fetal bovine serum and 100 U/ml penicillin/streptomycin solution at 5% CO ₂ and 37°C for 24 hours. The composition of the example was prepared with a concentration of 0 to 10%, and the cultured cells were treated with the composition for 6 hours. The cells were then treated with urban dust at a concentration of 200 μg/ml for 18 hours to measure cell viability. For measurement, after removing the culture medium, the cells were treated with a solution of MTS (0.33 mg/ml MTS) for a period of 1 to 4 hours, and the absorbance was measured at a wavelength of 490 nm using a spectrophotometer (Epoch 2, BioTek).

The results of the quantitative determination of MTS, the protective effect of human fibroblasts against exposure to urban dust, are shown in FIG. 5. As a result, in terms of cell viability from urban dust exposure, the group treated with the composition of the present invention showed 8 to 13% higher cell viability compared to the group not treated with the composition from urban dust.

Thus, it was confirmed that the present invention has an anti-pollution effect on the skin when exposed to air pollutants.

Example 4: Protection of cells from air pollutants and UV radiation (in vivo)

To confirm the cytoprotective effect of the composition of the present invention against exposure to air pollutants and ultraviolet radiation, human fibroblasts (NHDFs) were treated with urban dust as an air pollutant and irradiated with UV rays (UV-A) to evaluate the cytoprotective effect of the peptide composition. Cell viability was assessed by determining MTS ((3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazole).

Human fibroblast cells (normal human foreskin-derived dermal fibroblasts, NHDFs; ATCC, CRL-2522) were placed in a 24-well plate at a concentration of 3×10 ⁴ cells/well and cultured in DMEM containing 10% fetal bovine serum and 100 U/ml penicillin/streptomycin solution at 5% CO ₂ and 37°C for 24 hours. A sample (example composition) was prepared at a concentration of 1%, and cultured cells were treated with the composition for 24 hours and then treated with urban dust at a concentration of 80 μg/ml for 24 hours and irradiated with UV rays (UV-A) at a dose of 10 J /cm2 to measure cell viability. For measurement, after removing the culture medium, the cells were treated with a solution of MTS (0.33 mg/ml MTS) for a period of 1 to 4 hours, and the absorbance was measured at a wavelength of 490 nm using a spectrophotometer (Epoch 2, BioTek).

The results of the quantitative determination of MTS, the protective effect of human fibroblasts against exposure to urban dust (UV) and exposure to UV rays (UV-A) are presented in Fig. 6. As a result, the group treated with the composition of the present invention showed 16% or more increased cell viability compared to the group not treated with the urban dust and UVA (UV-A) protection composition.

Thus, it was confirmed that the present invention has a cytoprotective effect on the skin while being exposed to air pollutants and UV irradiation.

Example 5: Cellular oxidative stress caused by exposure to air pollutants and UV rays

To confirm the protective effect of the composition according to the present invention in relation to cellular oxidative stress, the formation of free radicals (reactive oxygen species (ROS)) was measured.

Quantitative determination of H ₂ DCFDA (2',7'-dichloro-dihydro-fluorescein diacetate) was used to determine the formation of intracellular free radicals (RFK). The quantification of H ₂ DCFDA (2',7'-dichloro-dihydro-fluorescein diacetate (DCFDA)) is based on the principle of converting H ₂ DCFDA to its fluorescent product DCF (2',7'-dichloro-dihydro-fluorescein).

Human fibroblast cells (normal human foreskin-derived dermal fibroblasts, NHDFs; ATCC, CRL-2522) were placed in a 24-well plate at a concentration of 3×10 ⁴ cells/well and cultured in DMEM containing 10% fetal bovine serum and 100 U/ml penicillin/streptomycin solution at 5% CO ₂ and 37°C for 24 hours. A sample (example formulation) was prepared at a concentration of 1%, cultured cells were treated with the formulation for 24 hours and then DCFDA (Fisher Scientific, D399) was added at a concentration of 10 µmol/l. The cells were then cultured for 30 minutes.

DCFDA was used diluted in Hank's buffered saline (HBSS). After removing the DCFDA solution, cells were washed with HBSS solution 2 times, treated with urban dust at a concentration of 80 μg/ml for 24 hours, and irradiated with UV rays (UV-A) at a dose of 10 J/cm ² . Intracellular fluorescence intensity was measured using a fluorimeter (Victor3,1420 Multilabel counter, Perkin Elmer) with excitation at 485 nm and emission at 520 nm. The results are presented in Fig. 7. It was confirmed that the free radical scavenging ability was improved by about 73% or more in the sample-treated group compared to the non-sample-treated group.

Example 6 Wrinkle Smoothing Effect

To confirm the wrinkle-smoothing effect of the composition of the present invention, an injection containing the composition of the example was used, as shown in Table. 1.

Studies were conducted on healthy women aged 35 to 55 years. Composition for injection in a volume of 2 ml was injected into the skin three times with a two-week interval in order to confirm the effect of smoothing wrinkles. Wrinkle condition was assessed using photography and 3D analysis.

The results are shown in FIG. 8 - Fig. 10 showing a comparison of photos before and after injections in areas prone to wrinkles.

On FIG. 8 to 10 show that after administering the composition of the present invention three times at two weeks interval, it was confirmed that the wrinkle smoothing rate increased by 10 to 20%.

Based on the above results, it was confirmed that the present invention has no toxic effect on skin cells and is able to protect skin cells from exposure to harmful substances and light from the external environment.

The above descriptions are merely illustrative of the technical concept of the present invention, and modifications and variations may be made by those skilled in the art to which the present invention pertains without departing from the gist of the present invention. In addition, the embodiments disclosed in the present invention are not intended to limit the technical intent of the present invention, but to clarify the technical intent, and the scope of the technical intent of the present invention is not limited to these embodiments. The scope of protection of the present invention is defined by the appended claims, and all technical concepts within an equivalent scope are to be understood as being within the scope of the present invention.

Claims (45)

1. Cosmetic composition for protecting the skin from exposure to harmful substances, light, or a combination thereof, containing: peptides and amino acids, where the peptides include nicotinoyl tripeptide, pentapeptide, nicotinoyl hexapeptide, sh-octapeptide, tetracarboxymethyl hexanoyl dipeptide, hexacarboxymethyl dipeptide and biotinoyl hexapeptide-2 amide, and amino acids include arginine, asparagine, aspartic acid, glycine, histidine, hydroxyproline, isoleucine, leucine, methionine, phenylalanine, proline, serine, and valine. 2. Cosmetic composition according to claim. 1, additionally containing hyaluronic acid, where the molecular weight of hyaluronic acid is from 500 KDa to 2 MDa. 3. The cosmetic composition according to claim 1, wherein said harmful substances include one or more substances selected from the group consisting of yellow dust, fine dust, cigarette smoke, vehicle exhaust, heavy metals and air pollutants. 4. Cosmetic composition according to claim 1, where said composition is in the form of a filler. 5. Cosmetic composition according to claim 1, wherein said composition has antioxidant activity. 6. The cosmetic composition according to claim 1, wherein said composition inhibits the formation of cyclobutane pyrimidine dimers (CPD). 7. Cosmetic composition according to claim 1, wherein said composition contains non-crosslinked hyaluronic acid. 8. Cosmetic composition according to claim 1 for use in smoothing wrinkles or improving elasticity. 9. Cosmetic composition according to claim. 1, where the peptides contain, based on 100 mass parts of sh-octapeptide, from 1 to 50 mass parts of nicotinoyl tripeptide; from 10 to 150 mass parts of the pentapeptide; from 0.05 to 10 mass parts of nicotinoyl hexapeptide; from 5 to 70 mass parts of tetracarboxymethyl hexanoyl dipeptide; from 15 to 80 mass parts of hexacarboxymethyl dipeptide; And 5 to 70 parts by weight of biotinoyl hexapeptide-2 amide, and amino acids contain, per 100 mass parts of valine, from 80 to 180 mass parts of arginine; from 10 to 60 mass parts of asparagine; from 20 to 80 mass parts of aspartic acid; from 30 to 100 mass parts of glycine; from 50 to 150 mass parts of histidine; from 5 to 40 mass parts of hydroxyproline; from 40 to 120 mass parts of isoleucine; from 50 to 150 mass parts of leucine; from 5 to 40 mass parts of methionine; from 40 to 120 mass parts of phenylalanine; from 10 to 50 mass parts of proline; And from 20 to 80 mass parts of serine. 10. Cosmetic composition according to p. 1, additionally containing from 1 to 5% of the mass. hyaluronic acid based on the total weight of the composition. 11. Pharmaceutical composition for the prevention or treatment of diseases caused by exposure to harmful substances, light, or combinations thereof, containing peptides and amino acids, where the peptides include nicotinoyl tripeptide, pentapeptide, nicotinoyl hexapeptide, sh-octapeptide, tetracarboxymethyl hexanoyl dipeptide, hexacarboxymethyl dipeptide and biotinoyl hexapeptide-2 amide, and amino acids include arginine, asparagine, aspartic acid, glycine, histidine, hydroxyproline, isoleucine, leucine, methionine, phenylalanine, proline, serine and valine, and wherein said disease is one or more diseases selected from the group consisting of atopic dermatitis, contact dermatitis, seborrheic dermatitis, erythema, sunburn, photosensitivity, acne, dry skin, and psoriasis. 12. The pharmaceutical composition of claim 11, wherein said composition inhibits skin cell death caused by exposure to harmful substances, light, or a combination thereof. 13. A pharmaceutical composition according to claim 11, wherein said composition is in the form of a filler for injection. 14. Pharmaceutical composition according to claim 11 containing cross-linked hyaluronic acid. 15. A filler composition for protecting the skin from exposure to harmful substances, light, or a combination thereof, containing: peptides and amino acids, where the peptides include nicotinoyl tripeptide, pentapeptide, nicotinoyl hexapeptide, sh-octapeptide, tetracarboxymethyl hexanoyl dipeptide, hexacarboxymethyl dipeptide and biotinoyl hexapeptide-2 amide, and amino acids include arginine, asparagine, aspartic acid, glycine, histidine, hydroxyproline, isoleucine, leucine, methionine, phenylalanine, proline, serine, and valine. 16. A filler composition according to claim 15, wherein said composition is in the form of an injectable filler.

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