KR102619897B1 - Novel peptides and cosmetic compositions for enhancing skin barrier or inhibiting skin pruritus comprising the same - Google Patents

Abstract

The present invention provides a novel peptide and a cosmetic composition containing the same for strengthening the skin barrier or suppressing skin itching. The peptide promotes bonding between epidermal cells and has the activity of significantly suppressing the expression of cytokines that cause itching.

Description

Novel peptides and cosmetic compositions containing the same for enhancing skin barrier or inhibiting skin itching {Novel peptides and cosmetic compositions for enhancing skin barrier or inhibiting skin pruritus comprising the same}

The present invention provides a novel peptide and a cosmetic composition containing the same for strengthening the skin barrier or suppressing skin itching. The peptide promotes bonding between epidermal cells and has the activity of significantly suppressing the expression of cytokines that cause itching.

Human skin is composed of the epidermis and dermis, excluding the outermost layer of the skin (keratin), and below it is composed of the subcutaneous tissue (hyperdermis), which includes adipose tissue. The epidermal layer is mainly composed of keratinocytes, and the dermal layer is mainly composed of dermal fibroblasts. .

Functional cosmetic materials targeting wrinkle improvement and skin aging prevention mainly promote collagen protein expression in dermal fibroblasts, the main cells of the dermis layer of the skin, or inhibit proteolytic enzymes that decompose collagen, natural products, compounds, peptide materials, and dermal cells. These are proteins such as growth factors and cytokines that promote growth. However, it is known to be very difficult for most active materials with these effects to actually pass through the epidermis layer of the skin and reach the dermal layer.

The epidermal layer serves as a barrier to suppress the penetration of external harmful substances, ultraviolet rays, and chemicals and maintain moisture. The main cells that perform this function are epidermal cells, which are known to maintain the skin barrier and regulate the immune response in the tissue below the dermal layer by secreting inflammatory regulators (cytokines, chemokines, etc.). It is known that the skin barrier function of the epidermal layer is due to the bond between epidermal cells, that is, the expression and activation of junction proteins between cells. These adapter proteins not only serve as a primary physical defense against infection outside the cell, but also serve as an important defense barrier that protects dermal cells and suppresses imbalances in immune function caused by chemical stimuli inside and outside the cell. Adaptive proteins are usually divided into four groups: tight junctions, adherent junctions, gap junctions, and desmosomes. Among these, strong junction proteins (e.g., claudins) (claudins, occludins, ZO-1, 2, etc.) protect the cell barrier, which functions to prevent the movement of molecules, especially water-soluble substances, through the space between cells and to suppress cell permeability. It is known to be a major contributing factor. Therefore, materials that can promote bonding between epidermal cells and/or increase the expression of adhesion proteins are expected to be useful for suppressing skin aging or skin wrinkle formation through protecting and activating the skin barrier.

Itching is an unpleasant sensation that causes a desire to scratch or rub the skin, and is a common symptom caused by many skin diseases, internal diseases, and mental illnesses. To treat itching, it is best to remove the cause, but since it is often difficult to identify the specific cause, it is common to administer general treatment according to the symptoms. Representative itchy diseases include atopic dermatitis, eczema, hand eczema, herpes, and coin-shaped eczema. Itching is related to the regulation of inflammation and immune factors, and various cytokines such as interferon-gamma (IFN-γ) and interleukin-6 are known.

Korean Patent No. 1817244 relates to a novel peptide and a cosmetic composition containing the same for inhibiting skin aging or skin wrinkle formation. A cosmetic composition containing a peptide of a specific sequence for inhibiting skin wrinkle formation is disclosed. Registration Patent No. 1817250 relates to a novel peptide and a cosmetic composition containing the same for suppressing skin aging or skin wrinkle formation, and Korean Patent No. 953515, a cosmetic composition containing a peptide of another specific sequence for suppressing skin wrinkle formation. The issue relates to a composition for improving skin itching, and a cosmetic composition for improving skin itching containing as an active ingredient one or more dipeptides selected from the group consisting of alanyl-glutamine and glycyl-glutamine is disclosed.

However, the novel peptide presented in the present invention and a cosmetic composition containing the same for strengthening the skin barrier or suppressing skin itching have not yet been disclosed.

1. Korean Patent No. 1817244 2. Korean Patent No. 1817250 3. Korean Patent No. 953515

The purpose of the present invention is to provide a novel peptide and a cosmetic composition containing the same for strengthening the skin barrier or suppressing skin itching. The present invention was completed by discovering the effects of synthetic peptides on strengthening the skin barrier and suppressing itching factors.

In order to achieve the above object, the present invention provides a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

Additionally, the present invention provides a cosmetic composition for strengthening the skin barrier containing a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

According to one example of the present invention, the present invention provides a cosmetic composition for preventing skin aging comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

According to another example of the present invention, the present invention provides a cosmetic composition for inhibiting skin wrinkle formation comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

According to another example of the present invention, the present invention provides a cosmetic composition for suppressing itching comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

In the present invention, itching is caused by atopic dermatitis, eczema, hand eczema, pompholytic eczema, coin-like eczema, senile pruritus, contact dermatitis, urticaria, psoriasis, and ultraviolet rays, or by interferon-gamma (interferon-γ) or interleukin. It may be caused by -6 (interleukin-6).

Additionally, the present invention provides a pharmaceutical composition for suppressing itching comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

The peptide of SEQ ID NO: 1, which is a specific peptide fragment according to the present invention, promotes excellent bonding between epidermal cells, induces skin barrier strengthening, and presents the effect of suppressing itching-causing factors, thereby strengthening the skin barrier, inhibiting skin aging, and reducing skin wrinkles. It can be usefully used to suppress formation or itching.

Figure 1 shows the results obtained by evaluating the permeability of FITC-dextran (4 kDa) through cell culture on transwell plates to measure cell binding and permeation inhibition of human epidermal keratinocytes according to the present invention. Lane 1 is a group in which cultured cells were not treated with the peptide sample (Control 1), and Lane 2 is a group in which cultured cells were treated with the control peptide, the peptide of SEQ ID NO: 2 (Control 2). , Lane 3 is the group treated with the peptide of SEQ ID NO: 1.
Figure 2 evaluates the resistance between cells using the Millicell-Electrical Resistance System (ERS) through cell culture on transwell plates to measure the electrical resistance (Trans-Epithelial Electric Resistance, TEER) of human epidermal cells according to the present invention. This shows the results obtained. Lane 1 is a group in which cultured cells were not treated with the peptide sample (Control 1), and Lane 2 is a group in which cultured cells were treated with the control peptide, the peptide of SEQ ID NO: 2 (Control 2). , Lane 3 is the group treated with the peptide of SEQ ID NO: 1.
Figure 3 shows the results obtained by measuring the secretion of the cytokine interleukin-6 (IL-6) by IL-6 ELISA by treating human epidermal cells with interferon-gamma (IFN-γ) according to the present invention. It is shown. Lane 1 is a group in which cultured cells were not treated with the peptide sample (Control 1), and Lane 2 is a group in which cultured cells were treated with the control peptide, the peptide of SEQ ID NO: 2 (Control 2). , Lane 3 is the group treated with the peptide of SEQ ID NO: 1.

Hereinafter, preferred embodiments of the present invention will be described in detail. In addition, in the following description, many specific details, such as specific components, are shown, but this is provided to facilitate a more general understanding of the present invention, and it is known in the art that the present invention can be practiced without these specific details. It will be self-evident to those who have the knowledge. Additionally, in describing the present invention, if it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the gist of the present invention, the detailed description will be omitted.

The present invention provides a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

The present invention provides a cosmetic composition for strengthening the skin barrier containing a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

In this specification, “skin barrier strengthening” means strengthening the barrier function of the stratum corneum located in the outermost layer of the skin. The skin barrier is composed of dead keratinocytes and intercellular lipids, and plays a key function in skin health as a skin protective film that protects the skin from external stimuli and prevents moisture from evaporating from the skin. Additionally, keratinocytes create a skin barrier through differentiation and keratinization processes. The skin barrier function can be destroyed by aging or external factors, and damage to the skin barrier can reduce skin moisture and cause wrinkles.

According to one example of the present invention, the present invention provides a cosmetic composition for preventing skin aging comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

As used herein, “skin aging” refers to skin aging caused by intrinsic and external factors, and includes, for example, skin photoaging accompanied by skin wrinkle formation, preferably Includes skin photoaging caused by ultraviolet rays, which is accompanied by the formation of skin wrinkles.

According to another example of the present invention, the present invention provides a cosmetic composition for inhibiting skin wrinkle formation comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

As used herein, “skin wrinkle” refers to a wrinkle formation on skin.

According to another example of the present invention, the present invention provides a cosmetic composition for suppressing itching comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

Additionally, in the composition according to one embodiment of the present invention, the composition may include additional ingredients useful for improving skin, and the additional ingredients may include all ingredients known to be effective in improving skin, including compounds and natural products. there is.

The cosmetic composition of the present invention includes ointment for external use, cream, softening lotion, nourishing lotion, pack, essence, hair tonic, shampoo, rinse, hair conditioner, hair treatment, gel, skin lotion, skin softener, skin toner, astringent, lotion. , from the group consisting of milk lotion, moisture lotion, nutritional lotion, massage cream, nutritional cream, moisture cream, hand cream, foundation, nutritional essence, sunscreen, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser. It may have any selected formulation, but is not limited thereto. The composition of each of these formulations may contain various bases and additives necessary and appropriate for the formulation of the formulation, and the types and amounts of these components can be easily selected by those skilled in the art.

When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier ingredient. You can.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as the carrier ingredient. In particular, when the formulation is a spray, chlorofluorohydrocarbon and propane may be used as carrier ingredients. /May contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan.

When the formulation of the present invention is a suspension, the carrier ingredients include water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, and microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.

When the formulation of the present invention is a surfactant-containing cleansing agent, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, and fatty acid amide. Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linoline derivative, or ethoxylated glycerol fatty acid ester can be used.

The formulation of the present invention may additionally contain excipients including fluorescent substances, fungicides, hydrotropes-inducing substances, moisturizers, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreens, vitamins, plant extracts, etc.

The preferred dosage of the composition of the present invention varies depending on the subject's condition and weight, degree, form, administration route and period of the disease, but can be appropriately selected by a person skilled in the art. However, for a desirable effect, the composition of the present invention may contain 0.01 to 10% of the peptide, more preferably 0.01 to 4%, based on the total weight of the composition on a dry weight basis.

In the present invention, itching encompasses all forms of itching that occur on the skin as a mental or physical symptom, preferably atopic dermatitis, eczema, hand eczema, herpes, coin-shaped eczema, itching in the elderly, contact dermatitis, urticaria, It may be caused by psoriasis and ultraviolet rays, or may be caused by interferon-gamma or interleukin-6, but is not limited thereto.

Additionally, the present invention provides a pharmaceutical composition for suppressing itching comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

In the pharmaceutical composition according to one embodiment of the present invention, the composition may include additional ingredients effective in suppressing itching, and the additional components may include all ingredients known to be effective in suppressing itching, including compounds and natural products. You can.

The pharmaceutical composition of the present invention can preferably be formulated in the form of an external skin preparation according to a conventional method. This formulation can be performed by a method commonly performed in the pharmaceutical field, and can be preferably formulated according to each disease or ingredient using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA.

The advantages and features of the present invention and methods for achieving them will become clear with reference to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below and will be implemented in various different forms. The present embodiments are merely intended to ensure that the disclosure of the present invention is complete and that common knowledge in the technical field to which the present invention pertains is not limited. It is provided to fully inform those who have the scope of the invention, and the present invention is only defined by the scope of the claims.

<Example 1> Synthesis of peptides

The peptides of SEQ ID NOs: 1 and 2 in Table 1 below were synthesized by the FMOC solid-phase method using an automated synthesizer (PeptrEx-R48, Peptron, Daejeon, Korea). The synthesized peptide was purified and analyzed by reverse-phase HPLC (Prominence LC-20AB, Shimadzu, Japan) using a C18 analytical RP column (Shiseido capcell pak), and mass spectrometry (HP 1100 Series). Identification was performed using LC/MSD (Hewlett-Packard, Roseville, USA).

Peptide name ^* sequence number Amino acid sequence (sequence length) Coll-acd SEQ ID NO: 1 REFHRKYRIPA (11-mer) Grgdsp (control peptide) SEQ ID NO: 2 GRGDSP (6-mer)

*: Peptide name: Random name given for experimental convenience

<Example 2> Human Epidermal Keratinocytes Culture

Human-derived epidermal cells were purchased and used from Lonza (Switzerland), and 1% of antibiotics was added to epidermal cell-specific medium (Keratinocyte Growth Medium, Lonza), cultured for 48 hours in a 5% CO ₂ incubator at 37°C, and then incubated in the existing cell culture medium (Keratinocyte Growth Medium, Lonza). The medium was removed, washed with phosphate-buffered saline (PBS), and the same medium was added and further cultured for use in the experiment.

<Example 3> Evaluation of cell binding and permeation inhibition efficacy between epidermal cells

To evaluate the efficacy of binding and permeation inhibition between epidermal cells, epidermal cells cultured in transwell plates were treated with FITC-dextran (4 kDa) and their permeability was measured. 3 x 10 ⁴ epidermal cells cultured in Example 2 were placed in 100 μl of a medium dedicated for epidermal cell culture, transferred to a 6.5 mm Transwell (ThermoFisher) containing a 0.4 μm Pore Polycarbonate Membrane Insert, and cultured for 24 hours. Afterwards, the existing medium was removed, the peptide sample (300 nM) was treated, cultured for an additional 24 hours, and then washed with phosphate-buffered saline solution. FITC-dextran (4 kDa) dissolved in phosphate-buffered saline was treated here and reacted in a 5% CO ₂ incubator for 5 minutes. Then, the transwell was removed and the fluorescence transmitted downward was measured.

As shown in Figure 1, the permeability was suppressed in the control peptide-treated group compared to the group not treated with the peptide, but the intensity of transmitted fluorescence in the group treated with the SEQ ID NO: 1 peptide was significantly lower, indicating that the SEQ ID NO: 1 peptide binds between epidermal cells. It was confirmed to inhibit cell permeation through the promotion of .

<Example 4> Evaluation of skin barrier efficacy of epidermal cells by measuring cell electrical resistance

To evaluate the skin barrier efficacy of epidermal cells, electrical resistance (TEER) was measured using the Millicell-Electrical Resistance System (ERS). 3 x 10 ⁴ epidermal cells cultured in Example 2 were placed in 100 μl of a medium dedicated for epidermal cell culture, transferred to a 6.5 mm Transwell (ThermoFisher) containing 0.4 μm Pore Polycarbonate Membrane Insert, and cultured for 24 hours. Afterwards, the existing medium was removed, the peptide sample (300 nM) was treated, cultured for an additional 24 hours, and then washed with phosphate-buffered saline solution. Here, to measure electrical resistance, the electrical resistance between cells was measured using a Millicell ERS-2 Voltohmmeter (Merck Millipore) containing an Ag/AgCl-electrode.

As shown in Figure 2, the electrical resistance increased in the control peptide-treated group compared to the group that was not treated with the peptide, but the electrical resistance was higher in the SEQ ID NO: 1 peptide-treated group, and SEQ ID NO: 1 peptide was used in the skin between epidermal cells. It was confirmed to have a barrier effect.

<Example 5> Evaluation of the effect on IL-6 expression by IFN-γ stimulation in epidermal cells

1 x 10 ⁶ epidermal cells cultured in Example 2 were transferred to a 100 mm culture dish in a medium dedicated for epidermal cell culture and cultured for 24 hours. Afterwards, the existing medium was removed, the peptide sample (300 nM) was pretreated for 2 hours, and then treated with IFN-γ (5 ng/ml) and cultured for an additional 24 hours. To measure secreted IL-6, cell culture medium was collected, concentrated 6-fold, and secreted amount was measured using the IL-6 ELISA kit (R&D systems) according to the suggested protocol.

IL-6 secretion increased by more than 3-fold compared to the control group due to stimulation of epidermal cells with IFN-γ. As shown in Figure 3, the secretion of IL-6 increased by IFN-γ stimulation was partially reduced in the control peptide-treated group, but the amount of IL-6 secreted was further reduced in the group treated with SEQ ID NO: 1 peptide. showed.

<110> Cha Meditech <120> Novel peptides and cosmetic compositions for enhancing skin barrier or inhibiting skin pruritus comprising the same <130>M21-0000 <160> 2 <170> KoPatentIn 3.0 <210> 1 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Coll-acd <400> 1 Arg Glu Phe His Arg Lys Tyr Arg Ile Pro Ala 1 5 10 <210> 2 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Grgdsp <400> 2 Gly Arg Gly Asp Ser Pro 1 5

Claims (12)

Peptide consisting of the amino acid sequence of SEQ ID NO: 1 Cosmetic composition for strengthening the skin barrier containing a peptide consisting of the amino acid sequence of SEQ ID NO: 1 delete delete Cosmetic composition for suppressing itching containing a peptide consisting of the amino acid sequence of SEQ ID NO: 1 The cosmetic composition for suppressing itching according to claim 5, wherein the itching is caused by atopic dermatitis, eczema, hand eczema, herpes, coin-like eczema, senile pruritus, contact dermatitis, urticaria, psoriasis, and ultraviolet rays. The cosmetic composition for suppressing itching according to claim 5, wherein the itching is caused by interferon-gamma (interferon-γ). The cosmetic composition for suppressing itching according to claim 5, wherein the itching is caused by an increase in interleukin-6. Pharmaceutical composition for suppressing itching comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1 The pharmaceutical composition for suppressing itching according to claim 9, wherein the itching is caused by atopic dermatitis, eczema, hand eczema, pompholytic eczema, coin-shaped eczema, geriatric pruritus, contact dermatitis, urticaria, psoriasis, and ultraviolet rays. The pharmaceutical composition for suppressing itching according to claim 9, wherein the itching is caused by interferon-gamma (interferon-γ). The pharmaceutical composition for suppressing itching according to claim 9, wherein the itching is caused by an increase in interleukin-6.