KR101100129B1 - Cosmetic composition included neolitsea aciculata extraction - Google Patents

Abstract

The present invention relates to a cosmetic composition containing the sapwood as an active ingredient.

Neolitsea aciculata extract of the present invention is prepared by extracting any one of lower alcohols selected from methanol or ethanol, and then sequentially dividing them into hexane, ethyl acetate, butanol and water having different polarities. It is composed.

According to the present invention, there is provided a cosmetic composition containing the aspen as an active ingredient extracts of the sakura wood having wrinkle improvement, antioxidant activity and skin lightening effect.

Birdwood, Extract, Fraction, Whitening, Antioxidant, Wrinkle Improvement

Description

Cosmetic composition containing sapling as an active ingredient {COSMETIC COMPOSITION INCLUDED NEOLITSEA ACICULATA EXTRACTION}

The present invention relates to a cosmetic composition containing the sapwood as an active ingredient.

In order to solve the root cause of human aging, modern science is conducting its research in various ways, and as life gets better, there is increasing interest in preventing aging issued to external parts as well as internal aging of the body.

In particular, in the external aging studies of humans, many studies are being conducted using skin.

The aging of the skin is called aging, which is a change in skin tissue due to chronologic aging and the prolonged exposure of sunlight to the skin. There is.

As the skin ages, the skin gets wrinkles and the skin loses its elasticity.

The most important issues addressed in the study of skin aging are free radicals and reactive oxigen species.

They are naturally produced in vivo, but are caused by pollution, solar ultraviolet rays, chemical oxidants, and microorganisms.

The free electrons or free radicals generated at this time exert oxidative stress on the skin cells, and the material produced in this process is considered to be the cause of melanin and wrinkle formation.

These reactive oxygen species break down the skin's antioxidant defenses made of enzymatic and non-enzymatic antioxidants, oxidative modification of biomolecules, damage to the skin barrier, cuts of connective tissue collagen and hyaluronic acid, and wrinkles caused by abnormal crosslinking. Accelerates skin aging

On the other hand, pigmentation caused by melanin production or melanocyte growth can be suppressed in various ways.

In general, various pathways such as inhibition of tyrosinase activity, an important enzyme of melanin synthesis, decreased function of melanocytes, reduction of melanin through inhibition of automatic oxidation, and inhibition of inflammatory reactions such as erythema are available.

As a result, in order to prevent skin aging, it is necessary to suppress the generation of excess active oxygen species and to efficiently remove the generated free radicals, and to inhibit the enzymes that cause wrinkles on the skin and inhibit the activity of the enzymes that proceed with pigmentation. Need research

Thus, researches using plants in relation to skin aging and whitening have been conducted as follows.

Korean Patent Publication No. 10-0874115 (cosmetic composition containing thyme extract as an active ingredient) relates to a cosmetic composition containing a thyme extract having an antioxidant effect, skin anti-aging effect, skin wrinkle improvement effect, whitening effect It is open.

Korean Patent Publication No. 10-0863267 (composition for improving skin condition containing herbal extracts), including ginseng, Astragalus, licorice, black axis, green tea and aloe extract as an active ingredient, whitening effect, skin strength and wrinkle improvement effect Disclosed is a composition for improving skin having a composition.

As described above, studies on various plants have been conducted, but research on cosmetic compositions using Saeedok has not been conducted.

The present invention, in order to solve the above problems, free radical scavenging ability, elastase activity inhibitory effect, tyrosinase activity inhibitory effect and melanogenesis (melanogenesis) inhibitory effect ( Neolitsea aciculata It is an object to provide a cosmetic composition having a skin wrinkle improvement, antioxidant activity and skin whitening effect.

The present invention relates to a cosmetic composition containing the sapwood as an active ingredient.

The sapling tree extract of the present invention is prepared by extracting any one of lower alcohols selected from methanol or ethanol, and then dividing them sequentially by adding hexane, ethyl acetate, butanol, and water having different polarities.

When described in more detail with respect to the present invention.

The present invention relates to a cosmetic composition containing the sapwood as an active ingredient.

Neolitsea aciculata is native to Korea and grows in wetlands of Jeju, Ulleungdo, and southern part of Japan and Korea.

Saeedok is an evergreen tree, a camphor tree, whose leaves are regenerated and grow at the ends of branches. Oval-shaped oval. The edges are flat, pointed and glossy, and the back is white and hairless.

Also known as the white-backed wood, it often uses leaves, bark, and fruit as dry and digestive agents.

The inventors of the present invention study various Jeju native plants to find components for blocking oxidation, delaying aging caused by oxidation, and inhibiting pigmentation of skin using Jeju Island native plants, which are clean areas. As a result, it was confirmed that the whitening effect as well as excellent antioxidant activity, wrinkle improvement and elasticity enhancing effect in Saeedok with the above effects.

Sadeokyi wood extract having such excellent activity can be applied as a functional cosmetic raw material, by completing the present invention by confirming that the use of natural plants can solve allergies and side effects, safety and stability caused by synthetic raw materials It was.

The present invention, the extract of the sakdeokgi, and confirmed the skin wrinkle improvement, antioxidant activity and skin whitening effect.

First, the present invention is repeatedly extracted 2 to 5 times by a method such as reflux extraction, shaking extraction or ethanol extraction using a solvent of about 10 to 40 times the volume of the weight of the saengduk tree branches, filtered and reduced the filtrate It is dried and prepared in powder form.

The extracts prepared in the form of powder are mixed with hexane, ethyl acetate, butanol, and water having different polarities, and sequentially fractionated. The extracts are dried under reduced pressure to prepare a powder form.

The present invention has been confirmed that the method of the method of extracting the Sapwood extract shows free radical scavenging activity using D PPH (1,1-Diphenyl-2-picrylhydrazyl radical) method, in particular ethyl acetate fraction It was confirmed that the extract and the butanol fraction extract showed high activity.

Free radicals are known as the causative agents of aging, especially skin aging.

The main cause of skin aging, particularly wrinkle formation, is the action of free radicals and the destruction of the extracellular matrix by matrix metalloproteinases (MMPs: collagenase, elastase, etc.).

Therefore, measurement of inhibitory activity of MMPs is very important for evaluation of skin aging inhibition.

Therefore, in the present invention, as a result of confirming the elastase inhibitory activity of the Edelase extracts, it was confirmed that the Edelase extracts reduced the elastase activity, in particular, high inhibitory activity in the ethyl acetate fraction extract It was confirmed.

In addition, since melanin is produced by tyrosinase acting as a key enzyme using L-tyrosine as a substrate, in the present invention, melanin measures tyrosinase activity inhibitory ability of the extracts of Pleurotus eryngii. It was.

As a result, it was confirmed that S. aureus extract reduced Tyrosinase activity, in particular, ethyl acetate fraction extract showed a high inhibitory activity.

In addition, as a result of measuring the melanogenesis inhibitory effect using the B16-F10 cell culture system in order to measure the whitening effect in the cell on the extract of Saengdeok tree, Saedok ethanol extract, hexane fraction extract, butanol fraction It was confirmed that the extract showed a high melanin synthesis inhibition rate.

This showed better activity than the inhibition rate of 1 mg / ml of arbutin used as a control, and the whitening activity in melanoma cells was better than that of arbutin.

In addition, as a result of confirming by the MTT method to confirm the cytotoxicity of the extracts of the present invention Saeduk tree, Saedok tree with less cytotoxicity to B16 melanoma cells and normal human fibroblast cells (NHF) It is thought that the development of functional cosmetic material with an extract as an additive is possible.

As described above, the saeedodeok extract of the present invention is excellent in antioxidant activity, wrinkle improvement effect, skin whitening effect can be very useful for food additives, beverage compositions, functional health foods, cosmetic compositions and the like.

In particular, when using the saengduk tree extract of the present invention as an active ingredient of the cosmetic composition, it contains 0.0001 to 10.0% by weight relative to the total weight of the cosmetic composition, preferably characterized in that it contains 0.1 to 10.0% by weight.

When the content of the extract is less than 0.0001% by weight, no wrinkle improvement, antioxidant activity and skin whitening effect are observed, and when the content of the extract is more than 10% by weight, the effect of increasing the content is insignificant, and there is a problem in the stability and stability of the formulation. It is not economical.

In addition, the cosmetic composition of the present invention may be prepared in any formulation that is conventionally prepared, and more specifically, may be prepared in the form of a lotion, essence, lotion, cream, pack, foundation, gel, ointment or spray. .

It may also be used as a pharmaceutical composition, may be prepared by methods known in the pharmaceutical art, and may be mixed with itself or with a pharmaceutically acceptable carrier, excipient, diluent, or the like to form a powder, granule, tablet, capsule or It may be prepared and used in the form of an injection or the like, and they may be administered orally or parenterally.

The effective dosage of the sapling extract according to the present invention may be appropriately selected depending on the absorbency, inactivation rate and excretion rate of the active ingredient in the body, the age, sex and condition of the patient, the severity of the disease to be treated.

According to the present invention, there is provided a cosmetic composition containing as an active ingredient the extract of Neophyllum ( Neolitsea aciculata ) having skin wrinkle improvement, antioxidant activity and skin lightening effect.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples, but these do not limit the scope of the present invention.

<Example 1> Preparation of Saeedopwood extract of the present invention 1

Neolitsea aciculata branches were prepared from Halla Arboretum.

After cutting 1 kg of the prepared saengdeok tree branch, it was put in 40 L of 80% ethanol and soaked for 1 week, and filtered.

The filtered filtrate was filtered and dried under reduced pressure to prepare a ethanol extract of sapling as a powder.

<Example 2> Preparation of Saeedopwood extract of the present invention 2

Neolitsea aciculata branches were prepared from Halla Arboretum.

After cutting 1 kg of the prepared saengdeok tree branch, it was put in 40 L of 80% ethanol and soaked for 1 week, and filtered.

The filtered filtrate was filtered, and hexane, ethyl acetate, butanol, and water having different polarities were added thereto, and the hexane fraction extract was dried under reduced pressure. The hexane fraction extract was prepared in powder form. .

<Example 3> Saeedok extract of the present invention 3

Prepared in the same manner as in Example 2, ethyl acetate fraction extract of the fraction extracts were dried under reduced pressure to prepare a ethyl acetate fraction extract of sapling wood powder.

<Example 4> Preparation of Saeedopwood extract of the present invention 4

Prepared in the same manner as in Example 2, the butanol fraction extract of the fraction extract was dried under reduced pressure to prepare a powdery sapling butanol fraction extract.

<Example 5> Preparation of Saeedopwood extract of the present invention 5

Prepared in the same manner as in Example 2, the water fraction extract of the fraction extract was dried under reduced pressure to prepare a water extract of Saeduk wood in powder form.

<Example 6> Preparation of the cosmetic composition containing the sapwood extract

The conventional cosmetic composition was used by adding 10% by weight of the cosmetic composition of Example 4 to the total weight of the cosmetic composition.

Experimental Example 1 Experiment of Determination of Free Radical Scavenging Activity by DPPH Method

1. Experimental Method

0.2 mM DPPH was dissolved in methanol for free radical scavenging activity, and each sample was added thereto, and then absorbance was measured at 517 nm with a spectrophotometer.

The inhibition rate was measured using the case where no sample was added as a control and the sample (extract of Example 1, 2, 3, 4, 5) as the experimental group.

Scavenging activity was expressed as the concentration of the sample (free radical scavenging activity (SC ₅₀ , μg / ml) required to reduce the DPPH concentration by 50%).

The free radical scavenging ability (%) is as follows.

= [1- (A-A ') / (B-B')] × 100

A: absorbance at 517 nm of the reaction solution to which the sample was added

A ': absorbance at 517 nm of the reaction solution without DPPH

B: absorbance at 517 nm of reaction solution without sample

B ': absorbance at 517 nm of sample and reaction solution without DPPH

2. Experimental results

As a result of the experiment, shown in Table 1 below.

SC ₅₀ (μg / ml) Example 1 (EtOH) 20.55 Example 2 (Hex) 161.1 Example 3 (EtOAc) 13.96 Example 4 (BuOH) 12.68 Example 5 (Water) 76.61

As shown in Table 1, the SC ₅₀ of the extract of Example 1 (ethanol extract) is 20.55 μg / ml, the extract of Example 2 (hexane fraction) is 161.1 μg / ml, the extract of Example 3 (ethyl acetate fraction) Silver 13.96 μg / ml, Example 4 extract (butanol fraction) was 12.68 μg / ml, Example 5 extract (water fraction) was found to be 76.61 μg / ml.

Thus, free radical scavenging activity was observed in all of the extracts of the present invention, and it was found that the extracts of Examples 3 and 4 exhibited high activity.

Experimental Example 2 Measurement Test of Elastase Inhibitory Activity of the Investigator Sae-duk-Tak Extract

1. Experimental Method

Each sample was added to 0.2 M Tris-Cl (pH 8.0) and 0.3 mM Elastase substrate SANA (N-succinyl- (Ala) 3-p-nitroanilide), followed by incubation at room temperature for 10 minutes. After preincubation, Elastase was added to measure the absorbance at 410 nm after reaction at room temperature for 10 minutes.

The control group was added to the solvent used as the sample solution instead of the sample, the experimental group was measured the inhibition rate as the extract of Examples 1, 2, 3, 4, 5 as the sample.

Elastase inhibitory activity was expressed as the concentration of the sample (IC ₅₀ , μg / ml) required to reduce the activity of the elastase by 50%.

Elastase inhibition rate (%) formula is as follows.

= [1- (enzyme activity of each sample) / (enzyme activity of control)] × 100

2. Experimental results

The experimental results are shown in Table 2 below.

IC ₅₀ (μg / ml) Example 1 (EtOH) 81 Example 2 (Hex) nd Example 3 (EtOAc) 119 Example 4 (BuOH) 195 Example 5 (Water) 1017

As shown in Table 2, as a result of measuring the elastase (Elastase) inhibitory activity of Saddockwood extracts (Examples 1,2,3,4,5), the extract of Example 1 (ethanol extract) IC ₅₀ was 81 μg / ml, the extract of Example 2 (hexane fraction) showed no activity, the extract of Example 3 (ethyl acetate fraction) was 119 μg / ml, the extract of Example 4 (butanol fraction) was 195 μg / ml, It was confirmed that the extract (water fraction) of Example 5 exhibited 1017 μg / ml.

According to the results, it was found that the elastase (elastase) inhibitory activity was the highest in the fraction extract of Example 3 of the Saeeduk wood fraction extracts (Examples 2, 3, 4, 5) of the present invention.

Experimental Example 3 Test of Tyrosinase Inhibitory Activity of the Extract of the Shingulus japonicus of the Present Invention

1. Experimental Method

0.1M potassium phosphate buffer (pH6.8), 0.2mM tyrosine, and 1250 units of tyrosinase were added for each concentration of the sample, followed by reaction at 37 ° C for 10 minutes, and then absorbance at 475 nm.

The inhibition rate was measured using the case where no sample was added as a control and the sample (extract of Example 1, 2, 3, 4, 5) as the experimental group.

Tyrosinase inhibitory activity was expressed as the concentration of the sample (IC ₅₀ , μg / ml) required to reduce the tyrosinase activity by 50%.

The tyrosinase inhibition rate (%) formula is as follows.

= [(A-B) / A] × 100

A: Absorbance at 475 nm of reaction solution without sample

B: absorbance at 475 nm of the reaction solution to which the sample was added

2. Experimental results

The experimental results are shown in Table 3 below.

IC ₅₀ (μg / ml) Example 1 (EtOH) 562 Example 2 (Hex) nd Example 3 (EtOAc) nd Example 4 (BuOH) 211 Example 5 (Water) 1121

As shown in Table 3, as a result of measuring the tyrosinase inhibitory activity that plays an important role in the production of melanin, in the case of the extract of Example 1 (ethanol extraction) IC ₅₀ is 562 μg / ml, the extract of Example 2 (hexane Fractions) and the extract of Example 3 (ethyl acetate fraction) could not confirm the mushroom tyrosinase inhibitory activity, the extract of Example 4 (butanol fraction) 211 μg / ml, the extract of Example 5 (water fraction) In the case of 1121 μg / ml.

Thus, the tyrosinase inhibitory activity was found to be the highest in the extract of Example 4 (butanol fraction).

<Experiment 4> Measurement experiment of melanin synthesis inhibitory effect of the present invention Saeeduk

1. Experimental Method

B16 melanoma cells were used to measure the inhibitory effect of melanogenesis.

Cells were aliquoted to 5 × 10 ⁴ cells / ml in 6 well plates and cultured for 24 h under 37% 5% CO ₂ conditions.

After washing with D-phosphate buffered saline (D-PBS) and exchanged with a medium containing 1 μM α-MSH, sample extracts (Examples 1,2,3,4,5) were added to each concentration and incubated for 3 days. .

After culturing for 3 days, the medium was removed, washed with D-PBS buffer, and then treated with trypsin to recover cells.

The recovered cells were added with 500 μl of 1M NaOH and left at 55 ° C. for 2 hours to obtain intracellular melnin.

This was measured for absorbance at 475nm with a microplate reader.

Arbutin was used as a control.

2. Experimental results

As a result of the experiment, it is shown in Figure 1 and Table 4 below.

Treatment concentration
(占 퐂 / ml) Melanin
% Biosynthesis inhibition rate Cell viability
(%) Control 100 0 100 Arbutin 1000 82 94 Example 1 (EtOH) 12.5
25
50
100 0
14
70.7
80 104
100
115
103 Example 2 (Hex) 12.5
25
50
100 0
80
88
92 91
79
70
62 Example 4 (BuOH) 12.5
25
50
100 14
35
44
81 101
102
107
80 Example 5 (water) 12.5
25
50
100 0
0
3
40 100
99
99
99

As shown in Table 4 and Figure 1, the inhibition rate of tyrosinase enzyme activity was confirmed, and the whitening effect was measured in cells using B16 melanoma cells.

The extract of Example 1 (ethanol extraction) showed an inhibition rate of 80% at 100 μg / ml, the extract of Example 2 (hexane fraction) of 100 μg / ml showed a 92% inhibition rate, the extract of Example 4 ( butanol fraction) showed melanin synthesis inhibition of 81%.

This showed a similar activity to that of 1000 μg / ml of arbutin (arbutin) used as a control, indicating that the whitening activity in melanoma cells was better than that of arbutin.

In addition, the cytotoxicity against B16 melanoma cells is low, and thus it may be possible to develop a cosmetic material related to whitening.

Experimental Example 5 Cytotoxicity Evaluation Experiment of M.D.

1. Experimental Method

In MTT assay, B16F10 melanoma cells and NHF (normal human fibroblaste) cells were incubated for 24 hours, and then sample extracts (Examples 1,2,3,4,5) were treated at various concentrations. 20 μl of 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H-tetrazolium bromide) was added per well.

After 4 hours of reaction at 37 ° C., the MTT solution was removed and 200 μl of DMSO was added per well.

After dissolving the formazan precipitate produced by reacting with living cells, the absorbance was measured at 540 nm with a microplate reader.

2. Experimental results

The experimental results are shown in Table 5 and FIG. 2 below.

Treatment concentration
(占 퐂 / ml) Cell viability
(%) Control 100 100 Example 1 (EtOH) 12.5
25
50
100 106
117
99
87 Example 2 (Hex) 12.5
25
50
100 88
64
52
31 Example 3 (EtOAc) 12.5
25
50
100 106
105
103
106 Example 4 (BuOH) 12.5
25
50
100 110
105
96
86 Example 5 (water) 12.5
25
50
100 99
100
100
94

As shown in Table 2 and Figure 2, as a result of confirming the cytotoxicity in the normal human fibroblast (NHF), in particular the extract (butanol fraction) of Example 4 showed excellent activity in B16 melanoma cells cytotoxicity It is thought that it will be useful for the development of functional cosmetic materials using it.

1 is a graph showing the melanin synthesis inhibitory effect in the B16-F10 cell culture system.

control: Control group (sample not added)

EtOH: Extract of Example 1

hex: Extract of Example 2

EtOAc: extract of Example 3

BuOH: Extract of Example 4

DW: Extract of Example 5

Figure 2 is a graph showing the cytotoxicity by the MTT method.

control: Control group (sample not added)

EtOH: Extract of Example 1

hex: Extract of Example 2

EtOAc: extract of Example 3

BuOH: Extract of Example 4

DW: Extract of Example 5

Claims (6)

As a cosmetic composition containing an extract of Neo-Della ( Neolitsea aciculata ) as an active ingredient, The sakura wood extract is characterized in that it has a skin wrinkle improvement, antioxidant activity and skin lightening effect, Cosmetic composition. The method of claim 1, The active ingredient is characterized in that it comprises 0.0001 ~ 10.0% by weight relative to the total weight of the cosmetic composition, Cosmetic composition. delete The method of claim 1, The sapwood extract is characterized in that it is prepared by extracting any one of lower alcohols selected from methanol or ethanol, Cosmetic composition. The method of claim 1, The sapling tree extract is extracted with any one of lower alcohols selected from methanol or ethanol, and then fractions are produced by sequentially dividing them into hexane, ethyl acetate, butanol, water of different polarity , Cosmetic composition. The method according to any one of claims 1, 2, 4 and 5, The cosmetic composition is characterized in that it has a formulation selected from the group consisting of lotion, essence, lotion, cream, pack, foundation, gel, ointment or spray, Cosmetic composition.

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