KR20050121397A - Asparagus cochinchinensis extract with irritation-palliative and anti-inflammatory effect and cosmetic composition for comprising same as active ingredient - Google Patents

Abstract

The present invention relates to an extract of astronomical dong having skin irritation and anti-inflammatory effect and a cosmetic composition containing the same. In particular, the astronomical extract of the present invention inhibits prostaglandin E ₂ production, which is an inflammation-inducing eicosanoid of the cycloclogenase pathway, and alleviates skin irritation caused by retinol and sodium lauryl sulfate, and thus, it is a lotion, essence, lotion, cream, It is available as cosmetics with formulations such as packs, gels, ointments, patches, or sprays.

Description

Astronomical extract and cosmetic composition containing the same

[TECHNICAL FIELD OF THE INVENTION]

The present invention relates to an extract of astronomical dong and skin cosmetic composition containing the same, having an anti-inflammatory effect and anti-inflammatory effect. _It relates to a cosmetic composition having an anti-inflammatory effect and a skin irritation-reducing effect that inhibits the production of ₂ .

[Private Technology]

The skin is a part directly exposed to the external environment, and when exposed to excessive ultraviolet rays or contaminants, skin irritation and inflammatory reactions such as erythema, edema, rash, tingling, and itching are induced. Skin troubles caused by these causes are not only aesthetic problems, but also substances produced during the inflammatory process may cause pigmentation of the skin and accelerate the breakdown of skin elastic fibers, thereby affecting the increase of skin wrinkles. It is known.

In external skin products used to protect the skin and for the cleansing of the skin, it is inevitable to use ingredients different from those for skin protection, such as surfactants, preservatives, fragrances, sunscreens, pigments, functional ingredients, etc. Are causing various side effects on the skin. In particular, ingredients such as AHA (alpha hydroxy acid), retinol, and arbutin contained in functional products for the purpose of whitening, wrinkle improvement, exfoliation, etc. are acting as the main cause of skin troubles. The substance is difficult to use at a concentration that can exhibit maximum efficacy.

For the same reason, researches on how to reduce the production of skin irritation and inflammation have been actively conducted. Kinins such as bradykinin activated by neuropeptides such as substance P secreted from neurons, or various cytokines formed by the COX pathway, PGE _2, etc. It has been shown to be involved in expansion and to cause inflammation and pain. However, neuropeptide antagonists, bradykinin antagonists, and COX inhibitors, including substance P, have been suggested as substances capable of reducing skin inflammation, but existing anti-inflammatory agents have problems in safety for skin or stability when containing products. Its use is limited.

In order to solve the problems of the prior art, an object of the present invention is to provide a substance that inhibits the production of PGE _{2 in} the COX pathway, inhibits inflammation, and alleviates skin irritation.

Another object of the present invention is to provide a composition for alleviating skin irritation or a composition for anti-inflammatory.

It is another object of the present invention to provide a cosmetic composition for skin irritation relief and anti-inflammatory.

In order to achieve the above object, the present invention provides an anti-inflammatory composition comprising the astronomical extract as an active ingredient.

In another aspect, the present invention provides a composition for alleviating skin irritation comprising the astronomical extract as an active ingredient.

In another aspect, the present invention provides a cosmetic composition having an anti-inflammatory activity or skin irritation-releasing activity comprising the astronomical extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present inventors studied plant extracts in order to alleviate skin irritation and search for substances having anti-inflammatory effects, and the astronomical extract extracts have an excellent effect of inhibiting the production of PGE ₂ , and sodium lauryl sulfate , Hereinafter referred to as "SDS") and retinol was found to be excellent in alleviating the skin irritation, based on this, the present invention was completed.

Astronomical dong according to the present invention is a scientific name, a tuber of Asparagus cochinchinensis ( Asparagus cochinchinensis ), a medicine that is harmless to the human body that has been used for medicinal purposes since ancient times.

The cheonmundong extract can be prepared by extracting or fractionating by adding water or an organic solvent in a weight ratio of 1: 1 to 1: 1000. The organic solvent may be an alcohol having 1 to 5 carbon atoms, hexane, acetone, ethyl acetate, methylene chloride or chloroform organic solvent, and the alcohol may be methanol, ethanol, butanol, isopropanol or dilution alcohol thereof, but is not limited thereto. . Preferably the organic solvent is ethanol, methanol or 30-99% dilute alcohol water thereof. The extraction method may be an immersion method or a layer separation used in the preparation of conventional plant extracts. For example, an alcohol crude extract is prepared, and then hexane, chloroform, ethyl acetate, and butanol are sequentially added thereto to separate the cheonmundong extract into layers. It can manufacture.

In one embodiment, 10 to 500 g of astronomical dong per 1 L of ethanol is added, followed by hot water extraction, preferably 60 to 120 ° C. for 1 to 24 hours, filtered and concentrated under reduced pressure to prepare an astronomical dong extract.

The astronomical extract according to the present invention inhibits prostaglandin production and has the effect of alleviating skin irritation by SLS and retinol. Thus, the astronomical dong extract is available for the purpose of anti-inflammatory and / or alleviating skin irritation.

Accordingly, the present invention provides an anti-inflammatory and / or skin irritation composition comprising the astronomical extract as an active ingredient. The anti-inflammatory and / or skin irritation-reducing composition may further include a pharmaceutically acceptable carrier, diluent or excipient in addition to the active ingredient, wherein the active ingredient is 0.0001 to 30% by weight of the total composition, preferably It may be included in 0.001 to 20% by weight, but is not limited thereto.

The carrier, excipient or diluent may be a substance added to conventional pharmaceuticals or cosmetics, such as glycerin, propylene glycol, hyaluronic acid, polyoxyethylene hardened oil, polyethylene, oleylethyl, ethanol, cetostearyl alcohol, Beeswax, dimethylsiloxane, sorbitan stearate, triethanolamine, monostearic acid, stearic acid, liquid paraffin, oils and betaines, which are appropriately selected depending on the formulation or method of use of the anti-inflammatory and / or skin irritant composition of the present invention. Use it.

In addition, the anti-inflammatory and / or skin irritation composition of the present invention may further comprise a preservative, fragrance, antibacterial, whitening agent, vitamins or pigments.

The anti-inflammatory and / or skin irritation composition according to the present invention can be used as a cosmetic, a medicament, a cleaning agent or a food additive, and preferably a cosmetic. When used as a cosmetic, the composition may be prepared in a formulation such as lotion, essence, cream, pack, gel, lotion, ointment, patch or spray.

Hereinafter, examples of the present invention will be described. The following examples are only for illustrating the present invention and the present invention is not limited to the following examples.

Example 1: Preparation of astronomical extract

50 g of dry weight of astronomical dong was put in 1 liter of ethanol, extracted by boiling at 80 to 100 ° C. for 12 hours in an extractor equipped with a cooling condenser, and filtered through a 300 mesh filter cloth. The filtrate was left at 4-15 ° C. for 7 days to mature, and then filtered through Whatman filter paper to prepare an astronomical extract .

Experimental Example 1: PGE of Astronomical Building ₂  Measurement of inhibitory efficacy

In order to verify the inhibitory effect of PGE ₂ production on astronomical dong, the following experiment was conducted (Ref .: Jeffrey, KH, Anglea, SW, Zoe, S., and Rhia CM Analytical Biochemistry , 1999, 271, 18-28).

Fibroblasts isolated from human epidermal tissue were passaged several times, and then 1 x 10 ⁵ cells were placed in 96-well plates and incubated for 24 hours in 37 ℃, 5% (v / v) CO ₂ incubator. After replacing with medium containing no FBS (fetal bovine serum) and incubated for 2 hours, the astronomical extract was treated to the concentration of Table 1, and then incubated for 2 hours at 37 ℃, 5% (v / v) CO ₂ incubator It was. Calcium ionophore A23187 and arachidonic acid were treated for 5 minutes at 50 uM concentration and dodecyltrimethylammonium bromide at a final concentration of 0.25% (w / v) for 10 minutes as a PGE ₂ stimulator. Cells were hemolyzed. 50 μl of the supernatant of the cell hemolyte was added to the plate coated with goat-anti mouse Ig, and the PGE ₂ standard samples were also prepared by concentration (20, 40, 80, 160, 240, 320, 480, 640 pg / 50). 50 μl each was added. 50 μl each of the PGE ₂ antibody and the PGE ₂ conjugate were added thereto, followed by reaction at room temperature for 2 hours using a plate stirrer. The medium and the buffer were removed, and washed four times. Thereafter, 150 µl of 3,3 ', 5,5'-tetramethylbenzidine (TMB)) / hydrogen peroxide (enzyme substrate) was added thereto and reacted with a stirrer for 30 minutes. Then, 1M sulfuric acid was added to stop the reaction. The reaction was quantified by measuring absorbance at 450 nm of ELISA reader to determine the effect of PGE ₂ inhibition of astronomical dong. The results are shown in Table 1 (5 repetition experiments) and FIG. 1. Inhibition rate of Table 1 is calculated based on the difference in the amount of PGE ₂ production of skin fibroblasts not treated with arachidonic acid and fibroblasts treated with arachidonic acid.

sample Concentration (ppm) PGE ₂ (pg / 10 ⁵ cells) Inhibition Rate (%) Negative Control (-Arachidonic Acid) - 26.2 ± 5.9 - Positive control (+ arachidonic acid) - 392.5 ± 11.4 - Astronomical Extract 10 282.8 ± 15.6 29.9 100 221.2 ± 20.8 46.8 1000 153.3 ± 18.4 65.3

In Table 1, the astronomical dong extract was observed to inhibit PGE ₂ production by arachidonic acid up to 65%, indicating that the astronomical dong extract inhibits the inflammatory response by inhibiting the induction or activity of the COX-2 enzyme.

Experimental Example 2: In vivo anti-inflammatory efficacy of astronomical extract

Animal experiments using astronomical extracts confirmed anti-inflammatory efficacy (mouse ear swelling test, MEST, Ref .: A. Crummey, GP Harper, EA Boyle and FR Mangan. Agents and Actions 1987, 20, 69-76).

Twenty-one hairless mice were divided into three groups of seven each, test group 1, which treated 0.5% (v / v) of astronomical extracts and 99.5% of ethanol as samples, 1.0% (v / v) of astronomical extracts + Test 2 treated as 99.0% ethanol as a sample and control group treated as% ethanol (v / v) as a sample.

Both ears of each group of mice were washed with ethanol prior to sample application, and then the average thickness of the ears of the mice was measured using a micrometer. Test group 1 and test group 2, 20 μl of each sample, and control group 20 μl of ethanol after 1 hour, ethanol in the left ear and arachidonic acid in the right ear of each mouse except the control 2 mg each was applied. After 1 hour of application, the thickness of both ears was repeated three times using a micrometer to measure the degree of edema of the ears by arachidonic acid. The left ear of the mice that were not coated with arachidonic acid was used as a control to determine whether inflammation occurred in Test Group 1 and Test Group 2, but no difference was observed as compared to before application of the sample. The anti-inflammatory effects of Test Group 1 and Test Group 2 were determined as edema inhibition based on the control group treated with ethanol, and the results are shown in Table 2.

Astronomical extract (% by volume) Ear thickness (㎛) % Inhibition Before arachidonic acid treatment After arachidonic acid treatment Thickness change Test group 1 0.5 319 526 207 30.3 Test group 2 1.0 327 514 187 37.0 Control - 321 618 297 - *% Inhibition = (AB) / A x 100A: average thickness change of control ear (thickness after arachidonic acid treatment-thickness before arachidonic acid treatment) B: average thickness change of test group ear (thickness after arachidonic acid treatment-arachidonic acid treatment Thickness before)

In Table 2, the test groups 1 and 2 treated with the astronomical dong extract inhibited the inflammatory response induced by arachidonic acid to 30.3% and 37.0%, respectively. Therefore, the anti-inflammatory effect of the astronomical extract was confirmed in the in vivo experiment.

Experimental Example 3 Measurement of Primary Stimulation and Stimulation Efficacy on Human Body

In order to investigate the safety of cheonmundong extract 1.0% (v / v) and the stimulating alleviation effect of cheonmundong extract on SLS, which is a stimulant, the human primary test was performed through the human patch test as follows. Stimulation tests were performed.

Fifty healthy men and women were tested according to CTFA guidelines (Ref .: The Cosmetic, Toility and Fragrance Association. Inc. Washington, DC, 20036, 1991). 15 μL of sample and solution in Finn Chamber. After dropping, it was placed on the back of the human body, which was the test site, and fixed with a tape.The patch was applied for 24 hours, the patch was removed, and after 4 hours, the skin reaction of the test site was determined according to the following criteria. The results are shown in Table 3 and FIG.

<Judge criteria>

0: no erythema or unusual symptoms

1: slightly redder than the surroundings

2: noticeably reddening around

3: more reddening and swelling than the surroundings

(formula)

Stimulus = ((0 x persons) + (1 x persons) + (2 x persons) + (3 x persons)) / number of subjects

sample Judgment result (person) Stimulus 3 2 One 0 Distilled water - 3 6 41 0.24 SLS (0.05% (w / v)) One 4 11 33 0.44 Astronomical extract (1% (v / v)) - 2 9 39 0.26 SLS (0.05% (w / v)) + astronomical extract (1% (v / v)) - 3 9 40 0.30

In Table 3, 1.0% (v / v) of astronomical dong extract was only about the same degree of stimulation as distilled water and significantly alleviated the stimulation by SLS 0.05% (w / v). These results are due to the excellent anti-inflammatory effect of the astronomical extract.

Experimental Example 4: PGE by Retinol Stimulation of Astronomical Aqueous Extract ₂  Measure generation inhibition

It was confirmed whether the astronomical dong extract can alleviate or inhibit stimulation caused by retinol.

Fibroblasts isolated from human epidermal tissue were passaged several times, and then 1 x 10 ⁵ cells were placed in 96-well plates for 24 hours. The culture medium was replaced with FBS-free medium for 2 hours, and the astronomical extract was treated at the concentrations shown in Table 4 and incubated for 1 hour. Retinol was treated here at the concentration of 1000 ppm, and it left to stand at 37 degreeC for 1 hour. After treatment with dodecyltrimethylammonium bromide at a final concentration of 0.25% (v / v) for 10 minutes, the cells were hemolyzed, and the supernatant was taken to inhibit the effect of PGE ₂ production by retinol stimulation of astronomical extract. (Enzyme-Linked Immunosorbent Assay, ELISA). The results are shown in Table 4 (five repeated experiments). In Table 4, the inhibition rate was calculated based on the difference in the amount of PGE ₂ production between the retinol-treated skin fibroblasts and the retinol-treated fibroblasts.

sample Astronomical extract (ppm) Retinol (ppm) PGE ₂ (pg / 10 ⁵ cells) Inhibition Rate (%) Control group 1 - - 27.9 ± 4.5 - Control 2 - 1000 209.4 ± 14.6 - Test group 1000 1000 125.3 ± 12.3 46.3

In Table 4, the astronomical dong extract inhibited PGE ₂ production increased by retinol stimulation to about 46.3%. That is, it was confirmed that the astronomical extract can effectively inhibit the skin inflammatory response caused by retinol.

Experimental Example 5: Verification of the safety of astronomical extract

5-1. Cytotoxicity

In order to verify the primary safety as a raw material used for external skin preparations, the cell death was confirmed by MTT method after culturing V79-4 cells (Chinese hamster, continuous cell line of lung tissue fibroblasts) in astronomical extract (Ref. Mossman T. Journal of Immunological Methods , 1983, 65, 55-63 ). Experimental results are shown in Table 5 as cytotoxicity (IC50 (Inhibitory Concentration 50): 50% of the cells killing the concentration).

sample IC ₅₀ (%, w / v) SLS 0.005 Astronomical Extract 5

As shown in Table 5, the IC ₅₀ value of the astronomical dong extract is 0.5, which is 1000 times lower than cytotoxicity compared to SLS, it can be seen that the cell safety is very excellent.

5-2. Allergy Assessment (LLNA)

As an experiment to confirm allergen induction by the astronomical dong extract, a local lymph node assay (LLNA) was performed (Reference: OECD GUIDELINE FOR THE TESTING OF CHEMICALS; DRAFT NEW GUIDELINE 429: Skin Sensitization; Local Lymph Node Assay).

After 20 days, 40% and 60% of the cheonmundong extract was applied to both ears of mouse (Balb / c) at 25 μl for 3 days, the auricular lymph nodes of the mice were isolated. The isolated lymph nodes were pulverized into single cells, incubated for 24 hours with the addition of radioisotope [ ³ H] -methylthymidine, and the amplification degree (dpm) of cell proliferation was measured. The results are shown in Table 6 below.

sample density(%) Lymph node count (cells / mL x 10 ⁷ ) dpm (average) S.I. (Amplitude) ethanol 60 1.92 1358 Transportation Astronomical Extract 20 2.01 1522 1.1 Astronomical Extract 40 1.88 1405 1.0 Astronomical Extract 60 2.15 1489 1.1 Disintegration per minute (dpm): The amount of [ ³ H] -methylthymidine inserted into the cell 2. SI (stimulation index): Sample's mean dpm divided by control's mean dpm

The allergy evaluation result is expressed as lymphocyte amplification of the sample group (Tanmun-dong extract treatment group) compared to the control group (ethanol) .In all concentrations, if the amplification degree (SI) of the sample is 3 or more and shows a tendency to increase in a concentration-dependent manner, Will be considered. However, in Table 6, the astronomical extract was confirmed that the amplification degree (S.I.) is 3 or less and there is no possibility of allergen.

From the above experiments, it can be seen that the astronomical extract has excellent anti-inflammatory and stimulating effects and excellent skin safety.

As described above, the astronomical extract of the present invention exhibits an excellent anti-inflammatory effect by inhibiting the production of PGE ₂ , an inflammation-inducing eicosanoid of the COX pathway, relieving skin irritation of stimulating factors such as SLS and retinol, and skin stability. This has a very high effect. Astronomical extract of the present invention can be provided as a cosmetic having excellent anti-inflammatory effect and skin irritation efficacy.

1 is a result of measuring the inhibition of production of prostaglandin E ₂ (hereinafter referred to as “PGE ₂ ”) at the cellular level of astronomical dong extract.

2 is a result of measuring the primary stimulation and stimulation relief effect of the astronomical extract of the human body.

Claims (8)

Anti-inflammatory composition comprising cheonmundong extract as an active ingredient. The composition of claim 1, wherein the astronomical dong extract inhibits prostaglandin production. The composition of claim 1, wherein the astronomical dong extract is extracted from astronomical dong with water or an organic solvent, and the organic solvent is an alcohol having 1 to 5 carbon atoms. A composition for alleviating skin irritation comprising cheonmundong extract as an active ingredient. The composition of claim 4, wherein the astronomical dong extract relieves skin irritation by retinol or sodium lauryl sulfate. The composition of claim 4, wherein the astronomical copper extract is extracted from astronomical copper with water or an organic solvent, and the organic solvent is an alcohol having 1 to 5 carbon atoms. Cosmetic composition having an anti-inflammatory activity or skin irritation activity comprising cheonmundong extract as an active ingredient. The composition of claim 7 wherein the formulation of the cosmetic composition is a lotion, essence, lotion, cream, pack, gel, ointment, patch or spray.