KR101452313B1 - Cosmetic composition having anti-inflammation and skin regeneration effect - Google Patents

Abstract

The present invention relates to a cosmetic composition having anti-inflammation and skin regenerating effect, which contains germinated barley extract as an active ingredient, and a cosmetic composition comprising a nucleic acid extracted from a salmon testis in order to double the effects of anti-inflammation and skin regen .

Description

[0001] Cosmetic composition having anti-inflammation and skin regeneration effect [0002]

The present invention relates to a cosmetic composition having anti-inflammation and skin regenerating effect, which contains germinated barley extract as an active ingredient, and a cosmetic composition comprising a nucleic acid extracted from a salmon testis in order to double the effects of anti-inflammation and skin regen .

Inflammation is one of the defense mechanisms that try to minimize or minimize the reaction when cells or tissues are damaged by causes such as stress or ultraviolet rays, causing cellular reactions with nerves, blood vessels, tissues, blood, etc. As a result, they may cause itching, fever, fever, pain, and tissue damage. Under appropriate conditions, normal function is restored after the initial state of inflammation. However, when stimulants stimulating the inflammation are not removed or are continuously produced, or when inflammation is induced by various chemical mediators or cytokines secreted by cells damaged by external stimuli, As enlargement occurs and permeability increases, antibodies, plasma and phagocyte cells are packed into inflammation sites, which causes skin inflammation and skin troubles.

Causes of inflammation include external factors such as external factors, immunological factors due to antigenic responses, immunological factors caused by cellular immune responses, physical factors due to climate, environment, and chemical factors. In addition, blood vessels and hormone imbalance .

Vaso-active amines, Baso-active polypeptides, and other mediators are the mediators responsible for inflammation. Examples of vasoactive amines include histamine and serotonin secreted from mast cells And it mainly acts to enhance vasodilation and permeability. In addition, the vasoactive polypeptides include kinin, plasmin, and complement, which act as vasoconstrictors. In addition, the vasoactive polypeptides include lymphokines such as interleukin-6 (IL-6) Acid (arachidonic acid) is responsible for the inflammatory reaction. The arachidonic acid produced by the phospholipase activated by the physical and chemical stimuli is then passed through the two pathways of cyclooxygenase (COX) or lipooxygenase to produce inflammatory mediators such as prostaglandin, leukotrienes and mediate various inflammatory responses (Thomas, R., Rev. Physiol. Biochem. Pharmacol., 100, 1984, published patent 10-2008-0020853).

In human keratinocytes and Langerhans cells, various cytokines can be produced by external substances and released. These cytokines are essential for inducing skin irritation and for progression of local inflammatory reactions (J Appl Toxicol 16: 65-70, 1996). In fact, in the case of irritant contact dermatitis, interleukin-6 (IL-6), interleukin-

1: IL-1) and Tumor Necrosis Factor-α (Contact Dermatitis 35: 355-60, 1996, Dec). In addition, there is a report that IL-6 and IL-6 (IL-6) are specifically increased by allergen-inducing substances and IL-6 and TNF-α are induced by applying specific chemicals (Am J Contact Dermat 8: 8: 158-64, 1997). The relationship of interleukin-8 (IL-8) to allergic reactions has also been reported (Contact Dermatitis, 35: 355-60, 1996 Dec). IL-1 and TNF-α are the cytokines involved in the stimulation, and IL-6 and IL-8 are allergen-related cytokines.

TNF-α is a typical cytokine secreted mainly by inflammatory cells or immune cells against external stimuli or infectious agents. It is associated with interleukin-1 (IL-1), inflammation, autoimmunity, allergy Is a proinflammatory cytokine that plays a central role in the mediation of sexually transmitted diseases. Chronic inflammatory diseases in the skin mediated by TNF-a include psoriasis, eczema or seborrheic dermatitis, and acute inflammatory diseases include contact dermatitis (Mache E and Descapms V, Ann. Dermatol. 1374-9, 2002). In addition, TNF-alpha-mediated allergic diseases include atopic dermatitis.

In order to inhibit TNF-a activity, a method of blocking TNF-α production, processing and secretion, or neutralizing TNF-α itself has been used. (TNF-α converting enzyme, TNF-α, TNF-α, TNF-α, TNF-α, (NIK, IKK, PKB) associated with the activity of NF-kB, a key signaling pathway leading to the expression of phosphodiesterase 4 and adenosine receptors, TNF-α, p38 and mitogen activated protein kinase (MAPK) such as c-jun N-terminal kinase (JNK) have been studied as a major target for inhibition of TNF-α (Patent Publication 10-2008-0051831).

Regulation of these cytokines and inhibition of the expression of inflammatory mediators can regulate stimulation and inflammation to normalize the skin. Removal of inflammation to eliminate inflammation, the action of reducing vital signs and symptoms is called anti-inflammatory. To date, substances used for antiinflammatory purposes are nonsteroidal system such as flufenamicacid, ibuprofen, benzydamine, indomethacin, and steroids such as prednisolone ), Dexamethasone, etc. In addition, neuropeptide antagonists, bradykinin antagonists, and COX inhibitors have been proposed. However, most of them have problems in terms of safety to skin and stability in cosmetic preparation, (Patent Publication No. 10-2008-0020853).

In order to solve the above problems, the inventors of the present invention have studied a combination of anti-inflammatory substances which can solve the problem of stability in the combination of anti-inflammatory effect, skin safety and cosmetic composition, and prescriptions for doubling the efficacy thereof. As a result, And that the effect of anti-inflammatory activity and skin regeneration component is doubled by further containing a nucleic acid extracted from a salmon testimony, thus completing the present invention.

Accordingly, it is an object of the present invention to provide a cosmetic composition that does not cause skin side effects, effectively inhibits skin inflammation, and has a skin cell regeneration effect.

The present invention provides a cosmetic composition having an anti-inflammatory effect and skin regenerating effect by further containing a nucleic acid extracted from a salmon testimony as an active ingredient in a germinated barley extract.

The germinated barley extract used in the present invention abundantly contains various active ingredients in addition to a substrate such as starch, and is widely used for many food materials. The germinated barley extract is known to contain a large amount of effective components such as γ-aminobutyric acid which has an effect of suppressing the rise of blood pressure, sedation of nerves and improvement of liver function, and glycine and betaine which are related to sweetness and sweetness .

The germinated barley extract is prepared by germinating barley, pulverizing it, extracting an active ingredient with an extraction solvent, and filtering the extract. Most preferably, water is used as an extraction solvent. Extraction is also possible by supercritical extraction.

The germinated barley extract is contained in an amount of 0.01 to 10% by weight based on the total weight of the composition. In the present invention, the germinated barley extract is contained in an amount of 0.01 to 10% by weight based on the total weight of the composition in order to obtain a desired effect. When it is used in an amount of less than 0.01% by weight, it is difficult to expect an appropriate effect. When it is used in an amount exceeding 10% by weight, the safety and stability of the product may be adversely affected.

It is also reported that the salmon testis-derived nucleic acid used in the present invention has cell regeneration, improvement of immune function, inhibition of lipid peroxidation, protamine effect on antimicrobial activity and electron donating ability against gram positive bacteria.

The nucleic acid extracted from the salmon testis is contained in an amount of 0.001 to 10% by weight based on the total weight of the composition. If the content of each component is less than 0.001 wt%, it is difficult to expect an appropriate efficacy. If the content exceeds 10 wt%, the safety and stability of the product may be adversely affected.

The cosmetic composition of the present invention prevents or alleviates skin irritation and inflammation, and provides the effect of relieving or normalizing sensitive skin.

The cosmetic composition according to the present invention is not particularly limited to the formulations and may be applied to various cosmetic compositions such as softening lotion, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing lotion, cleansing foam, It may be formulated as a pack, powder, body lotion, body cream, body oil, body essence, body cleanser, hair dye, hair tonic, scalp treatment, lotion, ointment, gel, cream, patch or spray.

The cosmetic composition having anti-inflammation and skin regeneration effect containing the germinated barley extract provided by the present invention protects the skin from external stimuli, and is safe not only in primary skin irritation but also as a skin irritant signaling substance or an inflammatory mediator It can be used in cosmetics for normalization of inflammatory skin and cosmetics for sensitive skin by regulating cytokines such as TNF-α and IL-6, 8 which are expressed.

1 is a photograph showing the skin regeneration effect on cells treated with Formulation Example 1 and Comparative Formulation Example 1 on human keratinocytes and normal cells not treated with a cosmetic composition.
FIG. 2 shows the results of analysis of IL-6 gene expression in human keratinocytes when Formulation Example 1 and Comparative Formulation Examples 1 and 2 were treated.
FIG. 3 shows the results of analysis of IL-8 gene expression in human keratinocytes when Formulation Example 1 and Comparative Formulation Examples 1 and 2 were treated.

Hereinafter, the composition and effect of the present invention will be described in more detail with reference to examples and test examples, but the present invention is not limited to these examples.

[Reference Example 1] Preparation of germinated barley extract

10 kg of barley was dipped in water at room temperature (12 ± 1 ° C) for 24 hours, germinated for 24 hours at 28 ° C in acacil. After the germinated barley was pulverized, 20 kg of purified water per 10 kg of the raw material was added to extract the active ingredient, and the extract was obtained using a 250 mesh net and a centrifuge.

[Reference Example 2] Preparation of nucleic acid derived from salmon testis

10 g of salmon testes were prepared and centrifuged at 1000 × g for 5 min. After discarding the supernatant, 10 ml of the first solution (50 mM glucose, 25 mM Tris, pH 8.0, 10 mM EDTA) Completely dissolved. To this was added 10 mL of a second solution (2 N NaOH, 1% sodium dodecyl sulfate), vortexed and held for 10 minutes, then phenol / chloroform / isoamyl alcohol (25: 24: And the organic solvent layer was mixed with the aqueous solution layer slowly and carefully at room temperature for 10 minutes. The mixture was centrifuged at 5,000xg for 15 minutes at room temperature to separate the aqueous solution layer and the organic solvent layer. At this time, the substance in the aqueous layer, which is like a thread, is the chromosomal DNA, which is extracted and dried at room temperature until the ethanol is blown, dried or connected with a vacuum pump, and nucleic acid derived from salmon testis is prepared.

[Reference Example 3] Preparation of Formulation Example 1 and Comparative Formulation Examples 1 and 2

The cream formulations in the form of milky lotions containing the germinated barley extracts obtained by the above-mentioned method were prepared as shown in Table 1 below.

Compounding ingredient Content (% by weight) Formulation Example 1 Comparative Formulation Example 1 Comparative Formulation Example 2 Germinated barley extract 1.00 1.00 - Nucleic acid derived from salmon testis 0.02 - - EDTA-2Na 0.02 0.02 0.02 glycerin 4.00 4.00 4.00 Butylene glycol 6.00 6.00 6.00 Pentaerythrityl tetraethyl hexanoate 3.00 3.00 3.00 Methofom seed oil 2.00 2.00 2.00 Squalane 2.00 2.00 2.00 Cetearyl alcohol 2.00 2.00 2.00 Behenyl alcohol 5.00 5.00 5.00 Phage-60 glyceryl isostearate 2.00 2.00 2.00 Chin type glyceryl stearate glucoside 1.00 1.00 1.00 Glyceryl stearate / phage-100 stearate 1.00 1.00 1.00 Hydrogenated Olive Oil Lauryl Ester 1.50 1.50 1.50 Sheer butter 2.50 2.50 2.50 Polyacrylate-13 / Polyisobutene / Polysorbate 20 0.30 0.30 0.30 Dimethicone 3.00 3.00 3.00 Preservative, fragrance, pigment Suitable amount Suitable amount Suitable amount Purified water Balance Balance Balance

[Test Example 1] Recovery and regeneration of skin cells: Cultivation of human keratinocytes

Human epidermal neonatal keratinocyte cells were cultured in order to examine the recovery and regeneration effect at the cell level. Human keratinocytes were purchased from Lonza, Inc. (Walkersville, MD, USA) and cultured in the CO ₂ incubator (CO ₂ incubator) at 37 ° C , And 5% CO ₂ . The cell culture was followed by the guidelines of Lonza, Inc. (Walkersville, MD, USA). The cells were suspended in 500 ml of KBM-2 (Clonetics CC-3103) medium using a KGM-2 bullet kit (bovine pituitary extract (BPE), human epidermal growth factor (hEGF) (Gentamycin Suflate + Amphofericin-B: GA-1) was added to the culture supernatant, and the concentration of insulin (0.5 ml), hydrocortisone (0.5 ml), transferrin (0.5 ml), epinephrine (0.5 ml) 1000, 0.5 ml)) was added thereto.

Human keratinocytes were cultured, and then Formulation Example 1 and Comparative Formulation Example 1 were added to human keratinocyte culture medium, respectively, and cells were cultured for 24 hours to prepare samples. On the other hand, as a control group, human keratinocytes not treated with a cosmetic composition were prepared. The cells of each sample were observed under an optical microscope, which is shown in Fig.

FIG. 1 shows that cells treated with Formulation Example 1 and Comparative Example 1, which are compositions containing germinated barley extract, recovered cells. In addition to the germinated barley extract, Formulation Example 1, which further contains nucleic acid extracted from salmon testes, showed remarkably superior cell regeneration effect than Comparative Formulation Example 1 containing germinated barley extract alone.

[Test Example 2] Inhibitory effect of IL-6 and IL-8 on expression: Quantitative-polymerase chain reaction (Q-PCR)

Human keratinocyte cultures were treated with interferon gamma after the cultivation of human keratinocytes in the same manner as in Test Example 1, and then Formulation Example 1 and Comparative Formulation Examples 1 and 2 were added, respectively. Cells were cultured for 24 hours to prepare samples Respectively. As a negative control group, normal cell groups without interferon gamma treatment were prepared, and human keratinocytes without a cosmetic composition treatment were prepared as a positive control group. Twenty four hours after the material treatment, cells were washed twice with 10 ml of phosphate buffered saline (PBS) and total RNA (total RNA) was extracted from the cells using a triazole (Trizol reagent, Invitrogen, Carlsbad, CA, USA) . The isolated RNA was further purified using a Qiagen RNA kit (Qiagen, Valencia, Calif.). CDNA was synthesized from the separated RNAs using a reverse transcription kit (Superscript Reverse Transcriptase (RT) II kit, Invitrogen, Carlsbad, Calif.) And amplified by real time reverse transcription The expression of IL-6 and IL-8 was quantitatively analyzed by polymerase chain reaction (Q-RT-PCR). The expression patterns of IL-6 (Hs00174131-m1) and IL-8 (Hs00174103_m1), which are inflammation inducers, were evaluated using a TaqMan gene expression system (TaqMan gene expression system, Applied Biosystems, Foster City, CA) 6 gene expression analysis result is shown in Fig. 2, and the result of IL-8 gene expression analysis is shown in Fig.

As can be seen in FIGS. 2 and 3, IL-6 and IL-8 gene expression was decreased when Comparative Formulation Example 1 and Formulation 1 containing germinated barley extract were treated. In addition, when Formulation 1 containing nucleic acid extracted from Salmon testimony in addition to germinated barley extract was treated, it was confirmed that the degree of suppression of IL-6 and IL-8 gene expression was much better than that of Comparative Example 1 have.

[Test Example 3] Evaluation of TNF-alpha biosynthesis inhibitory effect by stimulation with retinoic acid in dermal fibroblast

The skin fibroblasts isolated from human epidermal tissue were cultured in a 96-well test plate with DMEM (Dulbecco's Modified Eagle's Medium, BioWhitakker) containing 10% FBS (Fetal Bovine Serum, fetal bovine serum) ^Four wells were attached to each well and cultured for 24 hours. The cells were cultured in DMEM without FBS for 24 hours. Then, the germinated barley extracts and the salmon testis-derived nucleic acids prepared in Referential Examples 1 and 2 were treated with DMEM containing 1% FBS at the concentrations shown in Table 2 below Next, after culturing for 1 hour, the culture was taken, and the inhibitory effect of TNF-α on the biosynthesis of TNF-α was evaluated by performing ELISA (using Pharmingen, OptEIA Human TNF-α set) against 50 μl of each culture solution Respectively. As a negative control group, dermal fibroblasts without the test substance were used. The results are shown in Table 2 below.

ingredient Extract concentration (% by weight) TNF-a (pg / ml) % Inhibition rate * Negative control group - 1.12 - Retinoic acid (100 [mu] M) - 9.72 - Retinoic acid (100 μM) + germinated barley extract Germinated barley extract (1.0%) 5.38 50.4 Retinoic acid (100 μM) + germination barley extract + salmon testis-derived nucleic acid Germinated barley extract (1.0%), salmon testis-derived nucleic acid (0.02%), 2.10 88.6

* Calculated based on differences in TNF-α production from cells treated with retinoic acid-treated skin fibroblasts and retinoic acid.

From Table 2, it can be seen that the amount of TNF-α produced by treatment with retinoic acid, which is a stimulating substance, can be suppressed by treating a composition containing germinated barley extract. In particular, a composition containing germinated barley extract and nucleic acid extracted from salmon testis , The biosynthesis of TNF-α, which is an inflammatory mediator induced by retinoic acid stimulation, is significantly inhibited in dermal fibroblasts.

Claims (4)

Germinated barley extract and nucleic acid extracted from salmon testis,
The germinated barley extract is contained in an amount of 0.01 to 10% by weight based on the total weight of the cosmetic composition,
The nucleic acid extracted from the salmon testis is contained in an amount of 0.001 to 10% by weight based on the total weight of the cosmetic composition,
6, and interleukin-8 and inhibits biosynthesis of tumor necrosis factor-alpha (TNF-alpha) to inhibit skin inflammation.
A cosmetic composition for improving skin.
The method according to claim 1,
Characterized in that the germinated barley extract is contained in an amount of 1 wt% based on the total weight of the cosmetic composition, and the nucleic acid extracted from the salmon testis is contained in an amount of 0.02 wt% with respect to the total weight of the cosmetic composition.
A cosmetic composition for improving skin. delete delete

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