KR101517197B1 - Cosmetic composition comprising Brassica oleracea L. var. gemmifera Zenker extracts - Google Patents

Abstract

The present invention relates to a method for producing a callus extract, comprising the step of extracting callus as an active ingredient, wherein the callus extract is callus-derived, the callus extracting a callus from a hypocotyl of germinated drop cabbage seeds; And culturing the induced callus. The present invention also relates to a cosmetic composition comprising the extract of Cabbage sprout as an active ingredient.

Description

The present invention relates to a cosmetic composition comprising Brassica oleracea L. var. gemmifera Zenker extracts}

The present invention relates to a cosmetic composition comprising an extract of Cabbage Bark as an active ingredient. More particularly, the present invention relates to a cosmetic composition comprising, as an active ingredient, an extract of a drop cabbage comprising a callus extract, an organic solvent extract or a hot water extract.

Skin is an important organ that plays various roles such as protection function, barrier function, temperature control function, excretion function, respiration function. However, as the skin ages, its function is rapidly deteriorated to aging, the thickness of the skin becomes thin, the moisture content of the skin lowers, and the skin becomes dry and wrinkles, spots, freckles, skin elasticity decrease, pigmentation Resulting in various skin lesions. Especially, reactive oxygen radicals and free radicals, which are generated due to increase of ultraviolet ray amount, air pollution and excessive fatigue and stress of modern people, oxidize or denature biological components (proteins, nucleic acids, cell membrane lipids, etc.) .

Recently, there have been various researches and developments to overcome the skin aging phenomenon which is changed by various internal and external factors as the awareness of the environment and health is expanded. As a result, whitening, Conventional cosmetics developed variously to impart a cosmetic effect such as moisturizing, elasticity, and softening effect impart a cosmetic effect such as whitening, wrinkle prevention, moisturizing, elasticity, and softening effect to the skin alone or in combination, It is possible to produce side effects such as skin allergies and skin troubles by ignoring the constitutional characteristics of an individual and to manufacture them as uniform ingredients, and thus it is not suitable for the skin type or formation factors. Therefore, Satisfied with troublesome and uneconomical problems by purchasing natural cosmetics separately Might be able to effect the necessary requirements have been developed to be a cosmetic composition comprising a complex uses natural plants that can comprehensively meet the characteristics required extract as a functional cosmetic.

Accordingly, various cosmetics containing plant-derived useful ingredients have been developed. Plants have excellent totipotency, in which whole plants are reconstituted even when only a part of the leaves, stems, roots constituting the plants are transplanted. This typical performance is comparable to that of animal embryonic stem cells. Callus is induced when cells of mitotic tissue that divide cell in plant are cultured in nutrient medium. The callus is an oily tissue in which a wound cell restores the ability to divide and protects against scratches when a wound is injured in a plant, and a callus derived from the tissue of the plant is an amorphous form that has lost its ability to cause normal organ formation or tissue differentiation. As a tissue or cell mass, it is also known as the stem cell of a plant.

The callus can be induced in any part of plants such as leaves, stems, roots, rhizomes, and fruits through plant cell culture, particularly tissue culture, and is characterized in that it can be continuously produced by subculture. The basic principle of plant cell cultivation is that all plant cells use the biological fact that they have the ability to construct whole plants from which the cells are derived and that the induced callus has a positive effect on the protection and activation of dermal stem cells Can accept.

On the other hand, bell pepper cabbage is a bell pepper cabbage ( Brassica oleracea L. var . Also known as gemmifera Zenker , the English name is known as Brussels sprouts. The bell pepper cabbage has been cultivated in Brussels, Belgium since the 16th century, and it has been popularized in Europe since the 19th century. It has been cultivated in northern Europe and the United States including the UK.

Unlike the common cabbage, the stem of the cabbage grows long and the leaves are rarely attached to it. Stems grow to 60 ~ 90cm in height and 30 ~ 40 leaves in leaves. It has a diameter of 3 ~ 5cm and a mass of 10 ~ 15g. When cultivated for about two years, the flower buds come out from each cabbage and bloom and bear fruit. Like other cabbages, they like a cool climate, but the stem and the cabbage grow differently. When the stem grows, it requires a temperature of 18 to 20 ° C and high humidity, but it requires dry and relatively cool conditions (12 to 13 ° C) when the cabbage is free. Especially cold resistance is very strong.

The cabbage has excellent storage stability, and the content of nutrients such as protein, calcium, phosphorus, iron, vitamin A and vitamin C is much higher than that of ordinary cabbage.

In order to solve the above problems, the present inventors have developed a cosmetic composition that extracts active ingredients from callus extract, organic solvent extraction, or hot water extraction from drop cabbage, and has conducted clinical evaluation. As a result, Thus completing the present invention.

It is an object of the present invention to provide a cosmetic composition comprising an extract of a drop cabbage without cytotoxicity and skin side effects as an active ingredient.

Another object of the present invention is to provide a cosmetic composition comprising an extract of a dropwise cabbage having an excellent whitening effect and an antibacterial effect as an active ingredient.

It is still another object of the present invention to provide a cosmetic composition comprising an extract of Valium Chalice which is excellent in wrinkle removal, antioxidative effect and nitrogen oxide (NO) inhibitory effect as an active ingredient.

One aspect of the present invention relates to a cosmetic composition comprising an extract of bell pepper cabbage as an active ingredient. Wherein the cosmetic composition comprises a drop cabbage extract as an active ingredient and the drop cabbage extract is a callus extract, wherein the callus extraction comprises: inducing a callus from a hypocotyl of germinated drop cabbage seed; And culturing the induced calli.

The drop cabbage extract is characterized by containing 0.01 to 20% by weight based on the total weight of the cosmetic composition.

Another aspect of the present invention relates to a cosmetic comprising the cosmetic composition.

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The cosmetic composition comprising the extract of Cabbage extract of the present invention as an active ingredient is excellent in whitening effect, antibacterial effect, wrinkle removing effect, nitric oxide (NO) inhibiting effect and antioxidant effect and is free from cytotoxicity and skin side effects, Thus, it can be safely applied to a cosmetic composition.

FIG. 1 is a graph showing cytotoxicity of macrophylla of Cabbage extract according to one embodiment of the present invention. FIG.
FIG. 2 is a graph showing elastase inhibitory activity measured in relation to the wrinkle-reducing effect of the drop cabbage extract according to one embodiment of the present invention.
FIG. 3 is a graph showing the DPPH radical scavenging activity measured in relation to the antioxidative effect of the drop cabbage extract according to one embodiment of the present invention.
FIG. 4A is a photograph showing antibacterial effect of Staphylococcus aureus extract according to one embodiment of the present invention on Staphylococcus aureus.
FIG. 4b is a photograph showing the antibacterial effect of Candida albicans extract of the drop cabbage extract according to one embodiment of the present invention.
FIG. 5 is a graph showing the NO (nitric oxide) inhibitory effect of the drop cabbage extract according to one embodiment of the present invention.
FIG. 6 is a graph showing the results of extracellular melanin content test for testing the whitening effect of the drop cabbage extract according to one embodiment of the present invention. FIG.
FIG. 7 is a graph showing the results of an intracellular melanin content test for testing the whitening effect of the drop cabbage extract according to one embodiment of the present invention. FIG.

In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to be exemplary, self-explanatory, allowing for equivalent explanations of the present invention.

One aspect of the present invention relates to a cosmetic composition comprising an extract of bell pepper cabbage as an active ingredient. The cabbage extract according to the present invention does not show cytotoxicity and can exhibit a whitening effect, an antibacterial effect, an antioxidative effect and a wrinkle-reducing effect.

The drop cabbage extract used in the present invention can be used for callus extraction, organic solvent extraction or hot-water extraction of the drop cabbage.

At this time, in the present invention, all the parts of the drop cabbage may be used in the organic solvent extraction and hot water extraction.

In one embodiment, the organic solvent extraction may be performed by adding 10 to 100 parts by weight of water and 100 to 350 parts by weight of an organic solvent to 100 parts by weight of the filtrate. Under these conditions, useful components of the drop cabbage can be easily extracted without being damaged.

In this case, the filtrate may be one obtained by adding 50 to 200 parts by weight of an organic solvent to 100 parts by weight of drop cabbage, aging for 5 to 7 days, and then filtering. In this way, the active ingredient of the drop cabbage can be more effectively extracted during the filtration.

The filtration is carried out by a conventional method. For example, filter paper can be used, but the present invention is not limited thereto. In addition, the filtered filtrate can be further concentrated in a conventional manner. For example, the filtrate may be concentrated by adding it to a rotary vacuum concentrator, but it is not limited thereto.

 Examples of the organic solvent include acetone, ethyl acetate, diethyl ether, benzene, chloroform, hexane, and alcohols having 1 to 10 carbon atoms. These may be used alone or in combination of two or more. When such an organic solvent is used, the useful components of the cabbage can be easily extracted without being damaged.

The alcohols used in the organic solvent include, for example, lower monohydric alcohols such as methanol, ethanol, butanol, pentanol and hexanol, alcohols such as 1,2-pentanediol, 1,5-pentanediol, Heptanediol, octanediol, decanediol and the like, lower trivalent alcohols such as propylene glycol, 1,3-butylene glycol and glycerin, and the like can be used. These may be used alone or in combination of two or more.

In one embodiment, the organic solvent may be a mixture of hexane: butanol: ethyl acetate in a volume ratio of 1: 0.1 to 5: 0.1: 5. The drop cabbage may be extracted with an organic solvent using an organic solvent having the above mixing ratio, and then fractionated and concentrated under reduced pressure or dried to obtain the drop cabbage extract of the present invention. The active ingredient of the cabbage may be effectively extracted during the extraction.

In one embodiment, the hot water extraction can be performed by adding 100 to 500 parts by weight of water to 100 parts by weight of the drop cabbage and heating at 30 to 100 ° C for 1 to 10 hours. Preferably 60 ° C to 90 ° C for 1 to 10 hours. Under these conditions, useful components of the drop cabbage can be easily extracted without being damaged.

After the hot water extraction, extraction methods such as stirring extraction, reflux cooling extraction, cold extraction, ultrasonic extraction or supercritical extraction can be used. Preferably, the extract obtained after reflux cooling extraction is filtered, concentrated under reduced pressure or dried, Can be obtained.

In the present invention, seeds of the cabbage may be used for extracting the callus.

In one embodiment, the callus extraction comprises: (a) germinating the washed droplet cabbage seed; (b) deriving a callus from the hypocotyl of the germinated drop cabbage seed; And (c) culturing the induced calli.

In the step (a), the seeds of the drop cabbage were taken and added to a solution containing 1 ~ 2 drops of NaClO 3% and Tween 20, followed by shaking for 15 minutes, followed by washing in sterilized water for 3 to 5 minutes can do. The washed droplet cabbage seeds can be germinated by dipping on an induction medium. The induction medium can be used without limitation if there is a medium conventionally used for germination of plant seeds. For example, sucrose 30.0 to 40.0 g / L and agarose 5.0 to 10.0 g / L were added to MS medium (Murashinge and Skoog vitamin) solid medium or 1/2 MS vitamin solid medium But are not limited thereto.

The bivalve cabbage seeds may be cultured in the induction medium by maintaining the pH at 5.5 to 6.0 and the culture chamber temperature at 23 to 27 ° C. When the safflower seeds germinate, the seeds can be shielded from light for 3 to 5 days to induce hypodermic coating.

In the step (b), the hypocotyl of the painted cabbage may be introduced into the induction medium and the plant hormone may be added to induce the hypocotyl callus.

The induction medium may be the same induction medium as described above in step (a), and thus a description thereof will be omitted.

A plant hormone may be added to the induction medium to induce the callus. The plant hormone added may be one selected from the group consisting of indole-3-acetic acid (IAA), zeatin, NAA (naphthalene acetic acid), and benzylaminopurine (BA) or a mixture thereof. Preferably, the IAA and the zeatin may be mixed and used.

The plant hormones may be used in an amount of 0.1 to 2 mg / L, respectively. Preferably 0.5 to 1.5 mg / L, respectively. When the plant hormone is added to the induction medium under the above conditions, induction of adventitious root is suppressed to the utmost, and induction of the callus is most active.

In the step (c), the induced calli can be cultured in a large amount. The culture may be, for example, suspension culture, but is not limited thereto. Bulk cultivation of the callus induced during the culturing may be possible.

The cultured callus may be filtered, and the solids separated from the liquid component may be centrifuged to separate the residue, and the centrifuged supernatant may be used as a liquid phase, or may be dried and powdered.

The drop cabbage extract may contain 0.01 to 20% by weight based on the total weight of the cosmetic composition. Preferably 0.1 to 15% by weight. More preferably 2 to 10% by weight. The object of the present invention can be easily achieved in the above range.

The carrier, excipient or diluent which may be contained in the composition containing the drop cabbage extract may include purified water, oil, wax, fatty acid, fatty acid alcohol, fatty acid ester, surfactant, humectant, thickening agent, But are not limited to, viscosity stabilizers, chelating agents, buffers, lower alcohols, and the like.

Another aspect of the present invention relates to a cosmetic comprising as an active ingredient a cosmetic composition comprising the above-mentioned drop cabbage extract. The above-mentioned cosmetics may be formulated in accordance with a conventional method such as lotion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, wax foundation, pack, ointment and spray Can be used.

Hereinafter, the configuration and operation of the present invention will be described in more detail with reference to preferred embodiments of the present invention. However, the following examples are provided to aid understanding of the present invention, and the scope of the present invention is not limited to the following examples. The contents not described here are sufficiently technically inferior to those skilled in the art, and a description thereof will be omitted.

Example  1: Solvent extraction of bell pepper cabbage

2 kg of bell pepper cabbage was added to 2 kg of 70% ethanol, aged at room temperature for one week, and then filtered with a filter paper to obtain a filtrate. The filtrate was concentrated using a rotary vacuum evaporator (Eyela, Japan).

1 kg of the concentrated filtrate, 0.1 kg of water and 2 kg of an organic solvent were mixed. At this time, hexane, ethyl acetate and butanol were mixed and used as the organic solvent. The combined filtrate was concentrated under reduced pressure to obtain a dropwise cabbage solvent extract.

Example  2: bell pepper cabbage Hot water extraction

500 g of water was added to 100 g of drop cabbage, and the mixture was heated at 70 DEG C for 4 hours. The mixture was filtered, concentrated using a vacuum concentrator, and lyophilized to obtain a drop cabbage extract.

Example  3: bell pepper cabbage Callus extraction

50 g of the selected bell pepper cabbage seeds were added to a solution containing 1 to 2 drops of NaClO 3% and Tween 20, shaken for 15 minutes, and then washed in sterilized water for 5 minutes.

The washed safflower seeds were pelleted in a 1/2 MS vitamin solid medium supplemented with sucrose 30 g / L and agarose 7.5 g / L, pH was adjusted to 5.7, the culture chamber temperature was maintained at 23 to 27 ° C, Were cultured under an environment adjusted to 16day / 8night. The seeds of the cabbage sprouted germinated on day 3 of cultivation were subjected to cancer treatment for 3 days to induce a hypodermic coat (徒.).

The hypocotyls of the safflower cabbage seeds were applied to 1/2 MS vitamin solid medium supplemented with 30 g / L of sucrose and 7.5 g / L of agarose, and then 1.0 mg of indole-3-acetic acid (IAA) L and 1.0 mg / L zeatin were added to induce the hypocotyl callus, and the induced calli were suspended cultured.

The suspension-cultured callus was filtered with a cotton filter, the filtrate on the cotton filter was centrifuged, and the supernatant was taken to obtain a callus extract.

Experimental Example  1: Cytotoxicity test for extract of cabbage

To determine the degree of cytotoxicity of the extract of cabbage extract of the present invention against macrophages, the production of formazan, a coloring material in Tetrazolium salts, was measured by mitochondrial dehydrogenase. This is due to succinatetetrazolium reductase, a dehydrogenase present in intracellular mitochondrial electron transport system, which is metabolically active. It is effective only for surviving cells, and it is used that the color intensity is linearly correlated with the number of cells.

The mouse macrophages (RAW 264.7 cell line) were placed in RPMI 1640 medium containing 10% FBS and cultured for 24 hours. The macrophages were dispensed in 96-well plates at 1 × 10 ⁴ cells / well.

The extracts of Examples 1 to 3 were diluted to a concentration of 0.2 μg / ml, 0.1 ml was added to each well, and the cells were cultured in an incubator at 37 ° C. and 5% CO ₂ for 24 hours. After incubation, the supernatant of each well was taken separately, and 10 μl of assay solution (ez-cytox) was added to each well. After incubation for 2 hours in the incubator, the absorbance was measured at 450 nm using Enzyme-Linked Immunosorbent Assay (ELISA) Respectively. In this case, solvent (ethanol) was used as a negative control and 10% DMSO (dimethyl sulfoxide) was used as a positive control. The absorptivity of cell viability of the negative control group was calculated as 100% Respectively.

In FIG. 1, the drop cabbage extracts of Examples 1 to 3 did not show cytotoxicity.

Experimental Example  2: Wrinkle-improving effect test for bell pepper extract

In skin aging, especially wrinkle formation, action by active oxygen and destruction of extracellular matrix by elastase are considered to be the main causes. Therefore, the extract of Cabbage Cabbage according to the present invention was tested for the wrinkle-reducing effect by evaluating the inhibitory activity of the elastase.

A 1.6 mM elastase substrate (N-succinyl- (Ala) 3-p-nitroanilide) (SANA) and 0.2 mM Tris-HCl buffer (pH 8.0) were prepared. 0.165 ml of the above-mentioned Tris-HCl buffer, 0.005 ml of SANA, 0.01 ml of elastase and the drop cabbage extract of Examples 1 to 3 were diluted to a concentration of 0.2 / / ml in a 96-well plate, Minute, the absorbance was measured at 405 nm, and the elastase inhibition rate was calculated as described below, and the results are shown in Fig.

Elastase inhibition rate: ((control-sample) / control) * 100

Where control is the absorbance of the reaction solution to which no sample is added, and smaple is the absorbance of Examples 1 to 3.

In Fig. 2, the drop cabbage extracts of Examples 1 to 3 were found to be excellent in the effect of inhibiting elastase.

Experimental Example  3: Antioxidative effect test for extract of cabbage

The antioxidant activity of DPPH extract was evaluated by free radical scavenging ability.

DPPH (2,2-diphenyl-1-picrylhydrazyl) generates free radicals in ethanol. 0.5 ml of a 0.1 mM DPPH solution prepared from 0.4 ml of ethanol and 0.1 ml of each of the dilutions of Example 2 were diluted to a concentration of 0.125, 0.25, 0.5, and 1 占 퐂 / ml, respectively, to a final concentration of 10%. After stirring for 10 seconds at 4 ° C for 30 minutes, the absorbance was measured at 520 nm using an ELISA, and the free radical scavenging activity was calculated as described below, and the results are shown in FIG.

Free radical scavenging ability: 100- (sample / control) * 100

As a control, ascorbic acid (vitamin C) was used.

FIG. 3 shows that when the extract of Example 2 was tested at a concentration of 1 μg / ml, the free radical scavenging ability was 50%, which was superior to the free radical scavenging ability of the drop cabbage extract.

Experimental Example  4: Antibacterial effect test for extract of cabbage

The antibacterial effect of the drop cabbage extract obtained through Example 3 was tested.

As a microorganism, Staphyllococcus aureus ) and Candida albicans ( Candida albicans ) were inoculated into the medium at 3.0 × 10 ⁵ and cultured at 37 ° C. for 24 hours. TSA medium was used for the Staphylococcus aureus strain, and YM medium was used for the Candida albicans strain. Two paper disks were placed on the streaking medium, and 20 μl of each of the drop cabbage extracts of Examples 1 to 3 was inoculated in each case to observe the inhibition of the growth of the microorganisms. The growth inhibition was observed in FIGS. 4A and 4B Respectively.

4A and 4B, it was found that the drop cabbage extract of Example 3 exhibited an antibacterial effect against Staphylococcus aureus (Fig. 4A) and Candida albicans (Fig. 4B).

Experimental Example  5: Drops for cabbage extract Nitric oxide  Production inhibition test

The NO (nitric oxide) inhibitory effect was confirmed for the drop cabbage extract obtained through Example 3.

The mouse macrophages (RAW 264.7 cell line) were cultured in RPMI 1640 medium containing 10% FBS (FBS) for 24 hours. The cultured macrophages were cultured in 96-well plates at 1 × 10 ⁴ cells / well And LPS (Lipopolysaccharide) was added to each well at a concentration of 1.0 μg / ml.

The extracts of Examples 1 to 3 were diluted to a concentration of 0.2 μg / ml, 0.1 ml was added to each well, and the cells were cultured in an incubator at 37 ° C. and 5% CO ₂ for 24 hours. After incubation, only the supernatant of each well was taken separately, and each well was measured with a Nitric Oxide detection kit and shown in FIG. Distilled water was used as a positive control and LPS (Lipopolysaccharide) alone was used as a negative control.

In FIG. 5, the drop cabbage extracts of Examples 1 to 3 showed that LPS-induced NO was effectively inhibited.

Experimental Example  6: Whitening effect test on bell pepper extract

The melanogenesis inhibitory effect was tested by the method for testing the whitening effect of the drop cabbage extract of Examples 1 to 3.

Extra-cellular melanin content in the medium was measured in addition to the intra-cellular melanin content of mouse tumor cells (B16-BL6 cells) in order to more accurately measure melanin production inhibitory effect.

(1) Extra-cellular melanin content measurement

(B16-BL6 cells) were cultured for 24 hours, and then cultured in hy- droone (phenol red (-), α-MSH 50 nM) medium of hyclone at 37 ° C under 5% CO ₂ And then stimulated with the cultured B16-BL6 cells by treating with 50 nM melanocyte-stimulating hormone (a-melanocyte-stimulating hormone).

The stimulated B16-BL6 cells were treated with 25 μg / ml of each of the concentrations of Examples 1 to 3 according to the present invention. To compare the whitening effect, α-arbutin, which is known as a representative whitening substance, Example 1) was treated with 25 占 퐂 / ml and cultured. As a control group, only α-MSH was used.

After completion of the cultivation, the cells treated with Examples 1 to 3 and Comparative Example 1 were centrifuged to take the supernatant, and the absorbance of the supernatant was measured at 405 nm, and the results are shown in FIG.

(2) Measurement of intracellular melanin contents (intra-cellular melanin contents)

(B16-BL6 cells) were cultured for 24 hours, and then cultured in hy- droone (phenol red (-), α-MSH 50 nM) medium of hyclone at 37 ° C under 5% CO ₂ And then stimulated with the cultured B16-BL6 cells by treating with 50 nM melanocyte-stimulating hormone (a-melanocyte-stimulating hormone).

The stimulated B16-BL6 cells were treated with 25 μg / ml of each of the concentrations of Examples 1 to 3 according to the present invention. To compare the whitening effect, α-arbutin, which is known as a representative whitening substance, Example 1) was treated with 25 占 퐂 / ml and cultured. As a control group, only α-MSH was used.

1N NaOH solution containing 10% DMSO was treated at 80 ° C for 1 hour to dissolve melanin, and the supernatant was taken by centrifugation and the absorbance was measured at 405 nm The results are shown in FIG.

In FIGS. 6 and 7, the drop cabbage extracts of Examples 1 to 3 were found to be more effective than albutin of Comparative Example 1 in inhibiting melanogenesis, and thus being effective for skin whitening.

Claims (7)

The present invention relates to a method for producing an extract of cabbage,
The drop cabbage extract is callus-extracted,
Wherein the callus extraction comprises: deriving a callus from hypocotyl of germinated bell pepper cabbage seed; And
And culturing the induced callus. The cosmetic composition according to claim 1,
[Claim 2] The cosmetic composition according to claim 1, wherein the extract of the drop cabbage comprises 0.01 to 20% by weight based on the total weight of the cosmetic composition.
delete delete delete delete A cosmetic comprising the cosmetic composition of any one of claims 1 and 2.

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