KR101369238B1 - Composition for improving inflammatory skin condition including photosensitizer - Google Patents

Abstract

The present invention provides a composition for improving skin diseases comprising a peptide complex comprising the amino acid sequence Nicotinyl-GHK (Gly-His-Lys) as an active ingredient.
According to the present invention, it is effective in improving and treating inflammatory skin diseases such as acne, and can provide a composition for improving skin diseases having an effect effective in reducing free radicals in the body.

Description

Composition for improving inflammatory skin disease, including photosensitizers {COMPOSITION FOR IMPROVING INFLAMMATORY SKIN CONDITION INCLUDING PHOTOSENSITIZER}

The present invention relates to a composition for improving inflammatory skin diseases, including a photosensitizer, and more particularly, including specific peptide complexes and active ingredients, thereby removing the active oxygen in the skin and reducing skin inflammatory effects. A composition effective for the amelioration and treatment of a disease.

The skin of the human body is an indispensable organ for the maintenance of life, which physically and chemically protects the body from extraterrestrial and performs the biochemical functions necessary for metabolism of the whole body.

All abnormalities that appear on the skin, including human hair, hands and toenails, are called skin diseases, or skin diseases. Since the skin covers the surface of the body, there are many opportunities for direct contact with stimuli or various pathogens from the outside, and are greatly affected by the body. In addition, even minor changes of the skin can be seen by eye and touched by hand, and it is easy to perform pathological histological examination and examination of microorganisms by taking a part of the lesion, and to easily diagnose the diagnosis of individual diseases. The type is considerably larger than that of other organs, and the disease name is complicated. Relatively common skin diseases include atopic dermatitis, contact dermatitis, seborrheic dermatitis, urticaria, athlete's foot, psoriasis, psoriasis, herpes simplex, shingles, dry skin, housewives and acne.

On the other hand, due to aging of the cells, physiological natural phenomena, skin cells, which are the basic units that make up the skin, are not normally divided, and the regeneration period is gradually longer, and the skin loses its elasticity. It starts to occur. Currently, surgery or ointment and vitamin C therapy are used to repair skin wrinkles due to aging, but it is not enough to maintain a firm and elastic skin, which is a long-time desire of human beings. In addition, skin anti-aging and skin improving agents using natural products such as fruit acid or kimchi extract have been developed for many years. However, it only functions as a skin improving agent by inhibiting androgen, a male hormone, and thereby increasing sebum secretion. There is no effect as a treatment for the disease at all.

In addition, the cause of acne has not yet been identified, but the sebum is excessively produced by the male hormone, and such sebum is decomposed into free fatty acids and various low molecular substances by lipase produced by Propionibacterium acnes. It is known that inflammatory reactions occur due to stimulation by these free fatty acids and various low molecular weight substances. Acne is thus also a testosterone (androgen) -dependent chronic skin disease. The same is true for seborrheic dermatitis.

The inflammatory response is that arachidonic acid, a component of phospholipids in the skin's endothelium, is separated from cell membranes by phospholipase (PL) A ₂ , which is a cyclooxygenase. , COX) acts as a prostaglandin (PG) E ₂ , which plays an important role in the inflammatory response. And PLA ₂ is activated by free radicals . Acnes produces leukocytes and other free radicals.

COX includes COX-1 and COX-2, COX-1 is a constitutive enzyme, and COX-2 is an enzyme induced by inflammation during induction. In this way, the inflammatory response through the arachidonic acid and the COX-2 reaction is the main reaction of acne and dermatitis.

As a result, acne can lead to sebaceous overproduction, pores and keratinization . It is considered a disease caused by a combination of factors leading to the action of Acnes and inflammation.

Currently, various treatments are being made for acne caused by a combination of various factors. Representative treatment methods include exfoliants that dissolve keratin using strongly acidic substances, remove fatty free acids directly on acne, and apply antibiotics (Erythromycin, Clindamycin). , Benzoyl peroxide which has antibacterial action against acne bacteria, Vitamin A derivative which reduces the number of cotton cloths by inhibiting the formation of cotton cloth, etc., applied to the skin, taking oral antibiotics, using steroid drugs, laser Various methods, such as using a. However, these therapies are drugs and physical treatments that can cause various side effects when administered for a long time. In particular, antibiotics or steroid-based drugs can cause serious problems that build up or build up in the body.

Korean Patent Application No. KR10-2010-7013169 "External skin preparation for acne skin" Korean Patent Application No. KR10-2011-7013938 "1-amino-alkalinecyclohexane derivative for the treatment of inflammatory skin disease" Korean Patent Application No. KR10-2011-7021558 "Use of peptides and cosmetics or pharmaceutical compositions for the treatment and / or management of skin, mucous membranes and / or scalp"

The present invention has been made to solve the above problems, and an object thereof is to provide a composition effective for the improvement and treatment of inflammatory skin diseases such as acne.

Another object of the present invention to provide a composition for improving skin diseases having an effective effect on the reduction of free radicals in the body.

The present invention for achieving the above object, provides a composition for improving skin diseases comprising a peptide complex comprising the amino acid sequence Nicotinyl-GHK (Gly-His-Lys) as an active ingredient.

In addition, the complex is characterized in that it further comprises EGF (Epidermal Growth Factor), IGF (Insulin-like Grwoth Factor), SOD1 (Superoxide Dismutase 1) or a mixture thereof.

In addition, the content ratio of each complex in the complex of Nicotinyl-GHK (Gly-His-Lys) 5 to 20 parts by weight of EGF (Epidermal Growth Factor), IGF (Insulin-like Grwoth Factor) 5 to 20 parts by weight By weight, and 5 to 20 parts by weight of SOD1 (Superoxide Dismutase 1).

In addition, each content ratio in the complex is 10 parts by weight of EGF (Epidermal Growth Factor), 10 parts by weight of IGF (Insulin-like Grwoth Factor), and 100 parts by weight of the peptide of Nicotinyl-GHK (Gly-His-Lys). It is characterized in that 10 parts by weight of SOD1 (Superoxide Dismutase 1).

In addition, the composition is characterized in that it comprises amino levulinic acid (ALA) or methyl-amino levulinic acid (Methyl-ALA) substituted with a methyl group as an active ingredient.

In addition, the skin disease is characterized in that at least one selected from acne, atopy, urticaria, psoriasis, inflammatory diseases.

According to the present invention, it is effective in improving and treating inflammatory skin diseases such as acne, and can provide a composition for improving skin diseases having an effect effective in reducing free radicals in the body.

1 is a realtime PCR application graph according to an embodiment of the present invention.

The present invention provides a composition for improving skin diseases comprising a peptide complex comprising the amino acid sequence Nicotinyl-GHK (Gly-His-Lys) as an active ingredient.

Hereinafter, with reference to the accompanying drawings, the present invention will be described in detail.

The present invention relates to a composition effective for the improvement and treatment of skin diseases such as acne, atopic dermatitis, urticaria, psoriasis, inflammatory diseases and the like, and a composition comprising a peptide of a specific amino acid sequence and various additives.

Skin disease improvement composition according to the invention is characterized in that it comprises a peptide complex containing Nicotinyl-GHK (Gly-His-Lys) as an active ingredient.

In addition, the peptide complex may include a substance that helps to improve skin disease, and examples thereof include EGF (Epidermal Growth Factor), IGF (Insulin-like Grwoth Factor), and SOD1 (Superoxide Dismutase 1).

EGF (Epidermal Growth Factor) is a protein consisting of 53 amino acids as epidermal growth factor, induces cell division and promotes epidermal cell growth, and constituents of cell proliferation and dermis of epidermal and endothelial cells. It is a component that promotes the proliferation of fibroblasts that synthesizes collagen and has an excellent skin regeneration effect.

The IGF (Insulin-like Grwoth Factor) is an insulin-like growth factor, and there are IGF-I and IGF-II, and IGF-II is a protein that is structurally similar to IGF-I. IGF-I is a 7.6 kD, single-chain polypeptide hormone that is structurally similar to the proinsulin synthesized in liver and fibroblasts, indicating the action of growth hormone (GH).

When growth hormone (GH) is secreted from the pituitary gland, IGF is produced in the liver. IGF-1 secreted in this way is distributed throughout the body along the human body works. IGF is an extremely important factor for life support, whereas IGH is composed of 70 amino acids, whereas HGH is composed of 191 amino acids. IGF is another part of the growth hormone chain and its main action is most effective in relation to GH.

The SOD1 (Superoxide Dismutase 1) is an enzyme that reduces free radicals as an antioxidant enzyme. Free radicals are a by-product of metabolic processes in the body, and normally SOD enzymes first convert free radicals to hydrogen peroxide, which in turn acts as an enzyme called glutathione peroxidase (GPx) to convert it into toxic water. Therefore, when the SOD activity decreases, free radicals meet with nitrogen dioxide and become a highly toxic substance called ONOO-, which in turn attacks various cells of the body to promote aging and trigger the development of various diseases.

Each content ratio in the complex is 5 to 20 parts by weight of EGF (Epidermal Growth Factor) and 5 to 20 parts by weight of IGF (Insulin-like Grwoth Factor) based on 100 parts by weight of the peptide of Nicotinyl-GHK (Gly-His-Lys). Parts, and 5 to 20 parts by weight of Superoxide Dismutase 1 (SOD1), in particular 10 parts by weight of EGF (Epidermal Growth Factor), IGF (Insulin-) based on 100 parts by weight of the peptide of Nicotinyl-GHK (Gly-His-Lys). like Grwoth Factor) 10 parts by weight, and SOD1 (Superoxide Dismutase 1) 10 parts by weight is most preferred.

The composition for improving skin diseases according to the present invention may further include aminolevulinic acid (ALA) or methyl-aminolevulinic acid (Methyl-ALA) substituted with a methyl group as an active ingredient material together with the peptide complex. .

5-Aminolevulinic acid (ALA) is an aliphatic compound of C-5 and is one of the intermediates found in tetrapyrrole biosynthesis (porphyrin chlorophyll, heme, vitamin B12, etc.) of various organisms. Chlorophyll and It is an essential precursor to the biosynthesis process of tetraphyrrolle such as heme.

Methyl-ALA is one of the substituents of 5-ALA is substituted with a methyl group, by increasing the lipophilic of the hydrophilic 5-ALA to increase the skin permeability can increase the skin improvement and treatment effect.

Such skin diseases that can be improved from the composition comprising the peptide complex and the active ingredient according to the present invention include various diseases caused by skin aging and inflammation caused by free radicals in the living body, for example, acne, atopy, urticaria , Psoriasis and inflammatory diseases.

In a preferred embodiment for improving or treating skin diseases using the composition according to the present invention, it is preferable to use a combination of LED irradiation for the activation of the photosensitizer included as an active ingredient in the composition of the present invention.

In addition, in a more preferred embodiment of the present invention, the wavelength of the LED irradiation for the composition of the present invention is effective both short wavelength and long wavelength, most preferably short wavelength of 450 ~ 475 nm, long wavelength irradiation of 650 ~ 675 nm .

According to a more preferred embodiment of the present invention, the LED irradiation time is 5 to 20 minutes, even more preferably 5 to 15 minutes, most preferably 10 minutes.

According to a more preferred embodiment of the invention, the irradiation distance of the LED is within 10 cm, even more preferably within 5 cm, most preferably within 2 cm.

Hereinafter, the present invention will be described in more detail with reference to Examples. It should be understood, however, that these examples are for illustrative purposes only and are not to be construed as limiting the scope of the present invention.

[Example]

Peptides  Composite manufacturing

Superoxide Dismutase (SOD) 1 ppm,

Epidermal Growth Factor (EGF) 1 ppm,

Insulin-like Growth Factor (IGF) 1 ppm,

Vitamin Peptide 10ppm)

[Experimental Example]

1. Cell culture

An enzyme called cyclooxygenase (COX) exists in our body as two subtypes, COX-1 and COX-2. Among them, COX-1 is a protective enzyme found everywhere in the body, while COX-2 is mainly induced in an inflamed environment, and bacterial lipopolysaccharides, cytokines and growth factors. And expression is increased by tumor factors. In other words, COX-2 is an enzyme that promotes the synthesis of provocative prostaglandin.

In this experiment, the expression level of COX-2, an enzyme that causes inflammation in human keratinocytes (HaCaT), was determined at the mRNA level. , 10% Fatal bovine serum (FBS) and 1% Antibiotic-Antimycotic (GIBCO, Cat No. 15240-062) were incubated in a 75 Cm ² flask at 37 ℃, 5% CO ₂ conditions.

When the human keratinocytes were confluenced at 80% or more, 1 × 10 ⁵ cells / well were dispensed into 6well plates, and then cultured until cell confluence reached at least 80%. Thereafter, the medium was removed, and then, each sample was replaced with fresh medium, followed by further incubation under cell culture conditions for 24 hours.

The test method was real-time PCR, and the test procedure was as follows.

2. RNA Isolation

RNA was isolated by total RNA isolation using QiAzol Lysis Reagent (QIAGEN). The cells from which the medium has been removed are washed with PBS (phosphated buffer saline), and 1 ml of QIAzol Lysis Reagent is added and treated at room temperature for 10 minutes. After the transfer to a new E-tube 200ul chloroform (Amresco) was added and mixed well, and then reacted for 5 minutes at room temperature, centrifuged at 4 ℃, 12,000rpm for 15 minutes to separate the mRNA layer and protein DNA layer. 500 ul of the supernatant containing mRNA was transferred to a new E-tube, and 500 ul of Isopropanol (MERCK KGaA) was added thereto, inverted 4 to 5 times at room temperature, and further reacted at room temperature for 7 minutes. This was again centrifuged at 4 ℃, 13,000rpm for 30 minutes to precipitate the mRNA pellet. Remove the supernatant, wash the pellet with 1ml of 75% Ethanol, dry it, dissolve in 20ul of RNA Free Water (WelGENE Inc.), and use NanoDrop ^R Spectrophotometer (ND-1000, UV / VIS) to add 5ug of total RNA. Quantified by / ml.

3. cDNA synthesis

cDNA synthesis was synthesized using PrimeScript ^™ reagent kit (TaKaRa, RR037A). Specifically, cDNA was synthesized by mixing 50 μM oligo-dT primer 1 μl, 100 μM Random Primer 4 μl, 5X primeScript Buffer 4 μl, PrimeScript RT enzyme Mix 1 μl with 15 μL of total RNA, and reacting at 37 ° C. for 15 minutes to inactivate RTase. For 5 seconds at 85 ° C.

4. Real-time PCR

Real-time PCR was performed on the synthesized cDNA to analyze each gene. cDNA, a probe (beta-actin, COX-2), TaqMan Gene expression master mix (AB, Pro. No. 4369016) was mixed, and then performed using a 7500 Real time PCR system (applied biosystems). All genes were performed once at 50 ° C. for 2 minutes, at 95 ° C. for 10 minutes, and at 95 ° C. for 15 seconds at 60 times. Inventoried primers from Applied Biosystems (USA) were used. Expression rate was expressed by correcting the relative difference with each beta-actin.

[Experiment result]

In HaCaT, a human keratinocyte cell line, the effect of each treatment group of Table 1 on COX-2 expression was tested by real-time PCR (Table 1).

Light application conditions using the LED is shown in Table 2.

As a result, the group showing the highest effect of reducing COX-2 expression compared to the untreated control group was the treated group of Experimental Example 16. In particular, compared to the untreated control group, only red LED irradiation increased COX-2 expression and confirmed that it has an effect of inducing inflammation. Under such COX-2 stimulation conditions, ALA-2 and ALA-2 + P & P Complex It can be seen that there is an effect of reducing the expression of COX-2 (FIG. 1).

1 is a realtime PCR application graph according to an embodiment of the present invention.

division Test group The value of the difference
(relative amount of COX-2 expression relative to beta actin) Experimental Example 1 Control 1.00 Experimental Example 2 LED Blue only 1.09 Experimental Example 3 ALA-1 only 0.94 Experimental Example 4 ALA-2 only 1.04 Experimental Example 5 LED Blue + ALA-1 0.85 Experimental Example 6 LED Blue + ALA-2 1.01 Experimental Example 7 LED Blue + ALA-1 + P & P Complex 2.53 Experimental Example 8 LED Blue + ALA-2 + P & P Complex 4.43 Experimental Example 9 Control 1.0 Experimental Example 10 LED Red only 0.85 Experimental Example 11 ALA-1 only 1.82 Experimental Example 12 ALA-2 only 1.97 Experimental Example 13 LED Red + ALA-1 0.98 Experimental Example 14 LED Red + ALA-2 3.28 Experimental Example 15 LED Red + ALA-1 + P & P Complex 2.93 Experimental Example 16 LED Red + ALA-2 + P & P Complex 5.24

* P & P complex: peptide complex

ALA-1: 5-Aminolevulinic Acid Hydrochloride / CAS-No: 5451-09-2

ALA-2: 5-Aminolevulinic Acid Methyl Ester Hydrochloride / CAS-No: 79416-27-6

LED Blue: Short wavelength blue light

LED Red: Long wavelength red light

Control: Control

wavelength Set current Set voltage Street 450-475 nm (BLUE) 1.0-5.0A 5.0-15.0V 2 cm-10 cm 650-675 nm (RED) 1.0-5.0A 5.0-15.0V 2 cm-10 cm

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (6)

Amino acid sequence Nicotinyl-GHK (Gly-His-Lys); And
Epidermal Growth Factor (EGF), Insulin-like Grwoth Factor (IGF), Superoxide Dismutase 1 (SOD1), or mixtures thereof
Peptide complex comprising a; And
Active component of aminolevulinic acid (ALA) or methyl-aminolevulinic acid (Methyl-ALA) substituted with methyl group
Inflammatory skin disease improvement composition comprising as an active ingredient.
delete The method of claim 1,
The content ratio of each of the complexes is 5 to 20 parts by weight of EGF (Epidermal Growth Factor) and 5 to 20 parts by weight of IGF (Insulin-like Grwoth Factor) based on 100 parts by weight of the peptide of Nicotinyl-GHK (Gly-His-Lys). , And SOD1 (Superoxide Dismutase 1) 5 to 20 parts by weight of the composition for improving inflammatory skin disease, characterized in that.
The method of claim 3,
The content ratio of each complex in the complex is 10 parts by weight of EGF (Epidermal Growth Factor), 10 parts by weight of Insulin-like Grwoth Factor (IGF), and SOD1 (100 parts by weight of the peptide of Nicotinyl-GHK (Gly-His-Lys). Superoxide Dismutase 1) Composition for improving inflammatory skin disease, characterized in that 10 parts by weight.
delete delete

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