KR20140019536A - Cosmetic composition containing oreocnide fruticosa extraction - Google Patents

Abstract

The present invention relates to a cosmetic composition containing Oreocnide fruticosa extract as an active ingredient. The Oreocnide fruticosa extract in the present invention is produced by extracting with any one low grade alcohol selected from the methanol or ethanol, and then adding hexane, ethyl acetate, butanol, and water, which have different polarities, in consecutive order to fractionate it. By the present invention, the cosmetic composition containing the Oreocnide fruticosa extract, which is effective in anti-wrinkle, anti-oxidative activity, anti-inflammatory activity, and skin whitening, as the active ingredient.

Description

[0001] The present invention relates to a cosmetic composition containing oleic acid fruticosa extract,

The present invention relates to a cosmetic composition comprising an extract of Opuntia as an active ingredient.

In order to solve the fundamental cause of human aging, modern science is conducting research in various ways. As life becomes more and more rich, there is increasing interest to prevent aging which occurs in the external part as well as internal aging of the body. Especially, in the research on the external aging of human beings, many studies using skin have been carried out.

Chronologic aging in which the structure and function of the skin is continuously decreased with aging as the time elapses, and extrinsic aging (photoaging), which is a change in the skin tissue due to long-term exposure of the sun's rays have. This aging process causes the skin to dry out, wrinkles, and the skin to lose its elasticity.

Free radicals and reactive oxygen species are the most important treatments in skin aging research. They are naturally produced in vivo but are caused by pollution, sunlight, chemical oxidants and microorganisms. The free electrons and reactive oxygen species generated in this process induce oxidative stress on the skin cells, and the substances generated during this process are considered to be the causative agents of melanin and wrinkle formation. These active oxygen species disrupt skin antioxidant defense membranes composed of enzymatic and non-enzymatic antioxidants, oxidative deformation of biomolecules, damage of skin barrier and chain breaks such as collagen and hyaluronic acid which are connective tissues, and wrinkle formation by abnormal cross-linking Accelerate skin aging.

On the other hand, pigmentation caused by melanin production or melanocyte proliferation can be inhibited by various methods. In general, various pathways such as suppression of tyrosinase activity, an important enzyme of melanin synthesis, reduction of melanocyte function, reduction of melanin production through inhibition of autoxidation, and suppression of inflammatory reaction such as erythema can be used.

As a result, in order to prevent aging of the skin, it is necessary to inhibit the production of excess active oxygen species and to efficiently remove the generated active oxygen, to inhibit enzymes that cause skin wrinkles, Research is needed.

Korean Patent Registration No. 10-0874115 discloses a cosmetic composition containing an extract of thyme having an antioxidative effect, a skin aging-preventing effect, a skin wrinkle-improving effect, and a whitening effect.

Korean Patent Registration No. 10-0863267 discloses a composition for improving skin having a whitening effect, a skin elasticity and a wrinkle-reducing effect, which comprises ginseng, hwanggi, licorice, black shaft, green tea and aloe extract as an active ingredient.

Studies on various plants have been conducted as described above. However, research on the cosmetic compositions using non-sapwood has not yet been conducted.

Oreocnide fruticosa is native to Korea, native to Korea, Kyushu, and Shikoku in Japan. It is also known as rock boletus, which is a nettle and a deciduous shrub. The stem stands upright and grows about 2 m. The small branches are thin and dark purple, and the young branches have thin hairs. Leaves are alternate phyllotaxis, long elliptical shape with sawtooth on the edge, and the tip of the leaf is long like a tail. On the back side of the leaves, there are hairs at the time of childhood, white, with hairs on the front and back veins of the leaves. The flowers bloom in early spring and are a mature tree.

The inventors of the present invention conducted researches on various Jeju native plants in order to find out the ingredients for inhibiting oxidative action, delaying the aging action induced by oxidative action, and inhibiting pigmentation of skin by using native plants of Jeju Island, which is a clean area As a result, it was confirmed that there is an excellent antioxidant activity effect, wrinkle improvement and elasticity enhancement effect as well as a whitening effect in the above-mentioned Solanaceae. The present invention has been accomplished by confirming that it is possible to apply this extract as a raw material for functional cosmetics and solve allergy, side effects, safety and stability caused by synthetic raw materials by using natural plants .

Korean Patent Registration No. 10-0874115 Korean Patent Registration No. 10-0863267

The present invention is a free radical (free radical) scavenging, biyang tree extract having anti-inflammatory activity and melanin synthesis (melanogenesis) inhibition effect (Oreocnide antioxidant activity, anti-inflammatory activity, and skin whitening effect, including skin, hair, and fruticosa .

The present invention provides a cosmetic composition comprising an extract of Oreocnide fruticosa as an active ingredient.

The Oreocinide extract of the present invention fruticosa ) as an active ingredient has the effects of improving skin wrinkles, antioxidant activity, anti-inflammatory activity and skin whitening effect.

1 is a graph showing the inhibitory effect on melanin synthesis in the B16-F10 cell culture system.
2 is a graph showing cytotoxicity by the MTT method (normal human fibroblast).
Fig. 3 is a graph showing cytotoxicity by the MTT method (human keratinocyte cell).
FIG. 4 is a graph showing inhibitory effect on melanin synthesis in the RAW 264.7 cell culture system. FIG.

The present invention provides a cosmetic composition comprising an alopecific tree extract as an active ingredient.

The present invention extracts the above-mentioned Mannitol, and confirmed skin wrinkles, antioxidant activity, anti-inflammatory activity and skin whitening effect.

The present invention provides a method for preparing a Solanum tuberosus extract, which comprises extracting Solanum tuberosum with one of lower alcohols selected from methanol or ethanol, adding a solvent having a different polarity, and sequentially fractionating the solvent.

As the solvent having a polarity different from that of the lower alcohol, hexane, ethyl acetate, butanol, water and the like can be used, but it is not limited thereto.

The lower alcohol extraction step may be repeated 2-3 times using a solvent having a volume of about 20 to 30 times the weight of the tree trunks, filtered, and dried under reduced pressure to prepare a powder.

The present inventors have found that the above-mentioned extracts of the Propolis extract exhibit the scavenging activity of free radicals, which is a causative substance of skin aging using the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) Fraction extract and butanol fraction extract showed high activity.

In addition, in order to measure intracellular whitening effect on the Solanum tuberculosis extract, the melanogenesis inhibition effect of the B16-F10 cell culture system was measured, and it was confirmed that the inhibition rate of melanin synthesis was high in the Solanum tuberosum extract .

It was found that the inhibitory activity of arbutin at 1 mg / mL was better than that of arbutin, and that the whitening activity in melanoma cells was superior to that of arbutin.

In addition, the inhibitory effect of NO (nitric oxide) on the RAW 264.7 cell culture system was examined to determine the anti - inflammatory effect of the extract on the extract. Here, NO (nitric oxide) is a substance known to play an important role in inflammation induction.

In order to confirm the cytotoxicity of the extracts of Bombyx mori, the extracts of Bombyx mori were investigated by MTT method. As a result, B16 melanoma cells, normal human fibroblast cells (NHF) and human keratinocytes It was confirmed that there was almost no cytotoxicity to the cells.

INDUSTRIAL APPLICABILITY The extract of the present invention can be effectively used for food additives, beverage compositions, functional health foods, cosmetic compositions and the like because of its excellent antioxidant activity, skin wrinkle improving effect and skin whitening effect.

In particular, 0.0001 to 10.0% by weight, 0.001 to 10.0% by weight, or 0.1 to 10.0% by weight of the total composition of the cosmetic composition may be included in the cosmetic composition when it is used as an active ingredient of the cosmetic composition.

When the content of the extract is less than 0.0001% by weight, wrinkle improvement, antioxidative activity and skin whitening effect are not exhibited. When the content of the extract is 10% by weight or more, the effect of increasing the content of the extract is insignificant, And it is not economical.

In addition, the cosmetic composition of the present invention can be prepared in any formulations which are conventionally prepared, and more specifically, it can be formulated into lotions, essences, lotions, creams, packs, foundations, gels, ointments or sprays .

It may also be used as a pharmaceutical composition and may be prepared by known methods in the pharmaceutical field and may be prepared by mixing with itself or a pharmaceutically acceptable carrier, excipient, diluent or the like to give a powder, granule, tablet, Injections, and the like, and they can be administered orally or parenterally.

The effective dose of the mulberry tree extract according to the present invention can be appropriately selected according to the degree of absorption, inactivation rate and excretion rate of the active ingredient in the body, age, sex and condition of the patient, severity of the disease to be treated,

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> of the present invention Illiterate tree  Extract preparation

Oreocnide fruticosa ) were prepared and supplied by Halla Arboretum.

One kilogram of the prepared yolk was cut into small pieces and placed in 20 liters of 70% ethanol for one week, followed by filtration.

The filtrate was filtered and dried under reduced pressure to prepare a powder of a non-ethanolic ethanol extract.

< Example  2> of the present invention Illiterate tree  Extract preparation

Oreocnide fruticosa ) were prepared and supplied by Halla Arboretum.

One kilogram of the prepared yolk was cut into small pieces and placed in 20 liters of 70% ethanol for one week, followed by filtration.

The hexane, ethyl acetate, butanol, and water, which are different in polarity, were sequentially added to the filtered filtrate, and the hexane fraction extract was dried under reduced pressure to prepare a powder of the octane tree hexane fraction.

< Example  3> of the present invention Illiterate tree  Extract preparation

The ethyl acetate fraction extract in the fraction extract was dried in the same manner as in Example 2, and the fraction was extracted with ethyl acetate.

< Example  4> of the present invention Illiterate tree  Extract preparation

The butanol fraction extract of the fraction extract was prepared by the same method as in Example 2, and then dried under reduced pressure to prepare a powder of the non-canola bean fraction.

< Example  5> Illiterate tree  Extract preparation

The water fraction extract in the fraction extract was prepared by the same method as in Example 2, and then dried under reduced pressure to prepare a water fraction extract of Frecklewood.

< Example  6> Illiterate tree  Extracted Cosmetics  Preparation of the composition

The cosmetic composition of Examples 3 to 5 was added in an amount of 10% by weight based on the total weight of the cosmetic composition.

< Example  7> DPPH Freedom by law Radical  Measurement of scavenging activity

1. Experimental Method

For measurement of free radical scavenging activity, 0.2 mM DPPH was dissolved in methanol, each sample was added thereto, and the absorbance at 517 nm was measured with a microplate reader.

The inhibition rate was measured using the test sample in which no sample was added as a control group (extracts of Examples 1, 2, 3, 4 and 5) as experimental groups.

The scavenging activity was expressed as the concentration of free radical scavenging activity (SC ₅₀ , μg / mL) required to reduce the concentration of DPPH 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 addition of DPPH

B: Absorbance at 517 nm of the reaction solution to which no sample was added

B ': Absorbance at 517 nm of the reaction solution without addition of the sample and DPPH

2. Experimental results

The results of the experiment are shown in Table 1 below.

SC ₅₀ (μg / mL) Example 1 (EtOH) 45.7 Example 2 (Hex) 360.5 Example 3 (EtOAc) 24.8 Example 4 (BuOH) 28.0 Example 5 (Water) 122.8

As shown in Table 1, the SC ₅₀ of the extract (ethanol extract) of Example 1 was 45.7 μg / mL, the extract (hexane fraction) of Example 2 was 360.5 μg / mL, the extract of Example 3 (ethyl acetate fraction) , The extract (butanol fraction) of Example 4 was 28.0 μg / mL, and the extract (water fraction) of Example 5 was 122.8 μg / mL.

Thus, all of the extracts of the present invention showed free radical scavenging activity, and in particular, the extracts of Examples 3 and 4 showed high activity.

< Example  8> Illiterate tree  Experiments on the inhibitory effect of melanin synthesis on the extract

1. Experimental Method

Melanogenesis inhibitory effect of B16 melanoma cells was measured in order to measure intracellular whitening effect on the extract of.

Cells were seeded at 5 × 10 ⁴ cells / mL in a 6-well plate and cultured for 24 h at 37 ° C under 5% CO ₂ .

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

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

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

The absorbance was measured at 475 nm using a microplate reader.

As a control group, arbutin was used.

2. Experimental results

The results of the experiment are shown in FIG. 1 and Tables 2 and 3 below.

Treatment concentration
(占 퐂 / ml) Melanin
Biosynthesis inhibition rate (%) Cell survival rate
(%) Control 100 0 100 Arbutin 1000 68.2 103 Example 1 (EtOH) 100 30.6 68.1 Example 3 (EtOAc) 100 80.6 101.2 Example 4 (BuOH) 100 29.9 102.5 Example 5 (water) 100 14.8 104.3

Treatment concentration
(占 퐂 / ml) Melanin
Biosynthesis inhibition rate (%) Cell survival rate
(%) Control 100 0 100 Arbutin 1000 68.3 83.9 Example 3 (EtOAc) 25
50
100 -6.5
39.5
71.9 116.7
80.1
101.2 Example 4 (BuOH) 50
100
200 29.5
36.4
57.6 113.0
91.3
84.4

As shown in Tables 2 and 3 and FIG. 1, intracellular whitening effect was measured using B16 melanoma cells.

The extract of Example 1 (ethanol extract) showed an inhibition rate of 30.6% at 100 μg / ml, the extract of Example 2 (hexane fraction), the extract of Example 3 (ethyl acetate fraction) and the extract of Example 4 (Butanol fraction) showed inhibition rates of melanin synthesis of 97%, 80.6% and 29.9%, respectively.

It was shown that the activity of albumin was better than that of arbutin (1000 μg / ml), which was used as a control, indicating that whitening activity in Melanoma cells is superior to that of arbutin.

In addition, the extract of Example 3 (Ethyl acetate fraction) is thought to be able to be developed as a whitening-related cosmetic material because of its low cytotoxicity against B16 melanoma cells.

< Example  9> Illiterate tree  Evaluation of cytotoxicity by MTT method of extract

1. Experimental Method

B16F10 melanoma cells, normal human fibroblasts (NHF), human keratinocyte cells (HKC) were cultured for 24 hours, and then the sample extracts (Examples 1, 2, 3, 4 and 5) After the treatment, the cells were cultured for 2 days, and then 20 μl of 500 μg / ml MTT (3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H-tetrazolium bromide) was added. After reacting at 37 ° C for 4 hours, the MTT solution was removed and 200 μl of DMSO was added per well. The formazan precipitate formed by the reaction with living cells was dissolved and then the absorbance was measured at 540 nm with a microplate reader.

2. Experimental results

The experimental results are shown in Tables 2, 3, 4, 5 and 2 below.

Treatment concentration
(占 퐂 / ml) NHF cell survival rate (%) HKC survival rate
(%) Control 100 100 100 Example 1 (EtOH) 100 81.3 70.1 Example 2 (Hex) 100 77.5 62.9 Example 3 (EtOAc) 100 195.3 162.3 Example 4 (BuOH) 100 134.6 116.8 Example 5 (water) 100 123.1 121.9

Treatment concentration
(占 퐂 / ml) NHF cell survival rate (%) Control 100 100 Example 1 (EtOH) 25
50
100
200
300 114.5
89.8
98.4
78.6
100.1 Example 3 (EtOAc) 25
50
100
200
300 123.6
136.1
204.9
207.3
191.5

As shown in Tables 2, 3, 4 and 5 and FIGS. 1, 2 and 3, cytotoxicity of human fibroblast (NHF) and human keratinocyte (HKC) The extract of Example 3 (ethyl acetate fraction), which showed excellent activity in B16 melanoma cells, is not cytotoxic and may be useful for the development of functional cosmetic materials using the same in the future.

< Example  10> Illiterate tree  The inflammation inducer of the extract, NO ( Nitric oxide ) Inhibition effect measurement experiment

1. Experimental Method

Cells were seeded at a density of 3 × 10 ⁵ cells / mL in a 24-well plate and cultured for 18 h at 37 ° C in 5% CO _2. After the medium was removed, the cells were washed with D-phosphate buffered saline (D-PBS) After exchanging with medium containing 1 [mu] M LPS, sample extracts (Examples 1, 2, 3, 4, and 5) were added for each concentration and cultured for 1 day.

After culturing for 1 day, 100 μl of Griess reagent was added to 100 μl of culture medium to measure NO (nitric oxide), and 90 μl of MTT was added to the remaining medium to confirm cytotoxicity.

The absorbance was measured at 540 nm and 570 nm using a microplate reader.

2. Experimental results

The results of the above experiment are shown in Tables 6 and 7 and FIG.

Treatment concentration
(占 퐂 / ml) NO inhibition rate
(%) Cell survival rate
(%) Control 100 0 100 Example 1 (EtOH) 100 30.8 70.6 Example 2 (Hex) 100 51.1 75.3 Example 3 (EtOAc) 100 80.5 144.0 Example 4 (BuOH) 100 27.8 94.5 Example 5 (water) 100 10.6 100.5

Treatment concentration
(占 퐂 / ml) NO inhibition rate
(%) Cell survival rate
(%) Control 100 0 100 Example 3 (EtOAc) 12.5
25
50
100 -4.7
16.5
47.6
77.4 118.2
122.6
139.4
149.2

As shown in Tables 6 and 7 and FIG. 4, the inhibitory effect of NO (nitric oxide) on RAW 264.7 cells was confirmed. As a result, the NO inhibitory effect was excellent in the extract of Example 3 (ethyl acetate fraction).

Claims (6)

A cosmetic composition comprising an extract of Oreocnide fruticosa as an active ingredient. The method of claim 1,
The beech extract is a cosmetic composition, characterized in that it has a skin wrinkle improvement effect. The method of claim 1,
The cosmetic composition according to claim 1, wherein the extract has an antioxidative activity. The method of claim 1,
The cosmetic composition of claim 1, wherein the extract has a skin whitening effect. The method of claim 1,
The cosmetic composition according to claim 1, wherein the extract has a anti-inflammatory activity. The method of claim 1,
The cosmetic composition according to claim 1, wherein the composition comprises 0.0001 to 10.0% by weight of the total composition of the cosmetic composition.

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