KR20130051266A

KR20130051266A - Compositions for anti-aging

Info

Publication number: KR20130051266A
Application number: KR1020110116517A
Authority: KR
Inventors: 박녹현; 나용주; 박준성; 강영규; 배지현
Original assignee: (주)아모레퍼시픽
Priority date: 2011-11-09
Filing date: 2011-11-09
Publication date: 2013-05-20

Abstract

The present invention relates to a composition for anti-aging comprising at least one substance selected from the group consisting of pine needle extract and eye bean extract. The composition according to the present invention has anti-aging effects by reducing wrinkles and increasing skin elasticity, while maintaining the sunscreen efficacy of the epidermal cells, and having no photosensitive side effects, preventing and preventing aging, increasing skin elasticity, and skin moisture. It is useful for enhancing and promoting skin regeneration.

Description

Anti-aging composition {COMPOSITIONS for ANTI-AGING}

The present invention relates to an anti-aging composition.

Retinol is best known as a cosmetic for preventing and preventing skin aging. Retinol is converted into retinal and retinoic acid in vivo, and since some derivatives are found and synthesized in vivo, they are collectively referred to as retinoids. In particular, retinol promotes the differentiation of skin cells and promotes the biosynthesis of collagen (collagen fiber) that affects wrinkles and elastin (elastic fiber) that affects elasticity, thereby reducing wrinkles and increasing skin elasticity. It is known.

However, the retinoic acid converted after retinol absorption by the skin inhibits the expression of histidase (Histidase, L-histidine ammonia lyase (HAL)), thereby inhibiting the production of t-UCA (trans urocanic acid).

Accordingly, the present inventors have made diligent efforts to solve the above problems, the composition comprising at least one substance selected from the group consisting of pine needle extract and deciduous bean has an anti-aging effect, but does not inhibit the expression of histidase t It was confirmed that ultraviolet absorption by -UCA was enabled, and the present invention was completed.

Korean public application application 1996-028838

An object of the present invention is to provide a composition exhibiting an excellent anti-aging effect without photosensitive side effects.

In order to achieve the above object, the present invention provides a composition for anti-aging comprising at least one substance selected from the group consisting of pine needle extract and rat eye extract.

The composition according to the present invention can reduce wrinkles, increase skin elasticity, and have anti-aging effects, but do not have photosensitivity side effects that can maintain the sunscreen efficacy of the epidermal cells, anti-aging and prevention, increase skin elasticity, It is useful for enhancing skin moisture and promoting skin regeneration.

1 evaluates histidase gene expression (qPCR).
2 is the result of CPD ELISA.
3 shows the results of evaluating anti-aging efficacy (qPCR).

The present invention relates to a composition for anti-aging comprising at least one substance selected from the group consisting of pine needles extracts and rats eye extract.

In the anti-aging composition, which is an aspect of the present invention, the pine needle extract may be extracted from a tree belonging to the genus Pinus spruce. The tree belonging to the genus Pinus is Pinus densiflora ), White pine ( Pinus bungeana ), pine ( Pinus koraiensis ), pine tree ( Pinus parviflora , Gomsol ( Pinus thunbergii ), Rigi pine ( Pinus rigida ), pine wood ( Pinus banksiana ) or Strobe Pine ( Pinus strobus ) and the like, but is not limited thereto, for example, pine ( Pinus). densiflora ).

In the anti-aging composition of one aspect of the present invention, the rat bean has an effect of preventing aging, strengthening bones, and preventing cancer, and is called soybean sprout or bean sprout bean. For example, in one aspect of the present invention, the rat bean may be Rhynchosia nolubilis or Glycine max .

Rhynchosia , also known as medicinal beans Nolubilis ) is rich in protein and vitamins, and glycinein , an anticancer substance, shows antioxidant effects. Oriental medicine is known to have the effect of suppressing all the wind fever by letting down the vein is blocked. In addition, all poisons, including the poison of mineral medicines, and the blood circulation is active.

Glycine max , commonly known as black beans or black soybeans or black soybeans, contains high amounts of glycidane, anthocyanins and isoflavones. Glycidine has anti-cancer activity, and anthocyanin lowers cholesterol and suppresses arteriosclerosis. Isoflavones are supplemented with insufficient female hormone to maintain feminine body shape and stabilize body temperature and autonomic nerve. It is reported to be effective. In addition, since the antioxidant function to remove the active oxygen is more than four times higher than ordinary soybeans, it is known that the frost is also effective in preventing skin aging. That is, it is confirmed that the effect of enhancing the function of collagen in the skin to strengthen the elasticity of the skin, to give vitality to the skin, and to regenerate and heal skin cells.

The pine needle extract or red bean extract can be extracted by a conventional extraction method in the art, such as hot water extraction, organic solvent extraction or steam distillation can be used. In addition, in addition to extracts directly from pine needles or rats eye beans, extracts obtained from pine needles powder or rats eye powder can be applied without limitation.

In the anti-aging composition of one aspect of the present invention, the pine needle extract may be contained 0.001% to 40% based on the total weight of the composition. When the content of pine needle extract is less than 0.001%, it was observed that the anti-aging effect by the pine needle extract is not sufficient. On the other hand, when the content of the pine needle extract is greater than 40%, it was confirmed that the problem that the other active ingredients are not sufficiently contained. In view of the above, the pine needle extract may be contained in 0.005% to 35%, 0.01% to 30% or 0.05% to 25% with respect to the total weight of the composition.

In the anti-aging composition of one aspect of the present invention, the rat eye extract may be contained 0.001% to 50% based on the total weight of the composition. It was observed that the anti-aging effect by pine needle extract was insufficient when the content of rat bean extract was less than 0.001%. On the other hand, when the content of the rat eye extract is greater than 50%, it was confirmed that the problem is not contained enough other active ingredients. In view of the above, the rat bean extract may be contained in 0.005% to 45%, 0.01% to 40% or 0.05% to 35% relative to the total weight of the composition.

In the anti-aging composition, which is an aspect of the present invention, the weight ratio of the pine needle extract: rat bean extract may be 0.1 to 10: 1. When pine needle extract has a weight ratio less than 0.1 with respect to rat eye bean extract, it shows similar efficacy as rat eye bean extract, and when pine needle extract has a weight ratio of more than 10 with mouse eye extract, the pine needle fragrance is dark and the feeling is good. Did. In the above aspects, in the anti-aging composition of one aspect of the present invention, the weight ratio of pine needle extract: mouse eye extract is 0.3-8: 1, 0.5-6: 1, 0.7-4: 1 or 0.9-2: It may be 1, specifically 1: 1.

In the anti-aging composition of one aspect of the present invention, the composition may include those for wrinkle improvement. For example, in one aspect of the present invention, the composition may prevent damage and degeneration of collagen and elastic fibers in the dermis and promote production by promoting skin elasticity. In addition, the skin cycle of the epidermal cells can be returned to normal, and keratinocytes with excellent water retention can be maintained.

In addition, in the anti-aging composition of one aspect of the present invention, the composition may be one that does not have photosensitive side effects. For example, in the sense of the present invention, since the composition has an anti-aging effect similar to that of retinol and does not inhibit the expression of the histidase gene, normally expressed t-UCA can absorb ultraviolet rays, The side effect of sensitivity to photostimulation can be eliminated.

In the anti-aging composition of one aspect of the present invention, the composition may include a cosmetic composition.

On the other hand, in the anti-aging composition of one aspect of the present invention, the composition is softening longevity, astringent longevity, nourishing longevity, nutrition cream, massage cream, eye cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing It may include formulated with water, packs, makeup bases, foundations, body lotions, body creams, body oils, body essences, body cleansers, essences, soaps, emulsions, lotions, ointments, gels, creams, patches or sprays. .

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

Pine needle extract and Rat bean Preparation of extract

(1) Preparation of Pine Needle Extract

Pine needles ( Pinus densiflora ) was washed with purified water and dried, 200 g of granulated pine needle powder was added to 1 liter of an aqueous 70% ethanol solution, boiled for 12 hours in an extractor equipped with a cooling condenser, filtered with 300 mesh filter cloth, and filtered. After aging for 7 days at oC, it is filtered through Whatman No. 2 filter paper. The extract was added to a 3-liter separatory funnel, and 1 liter of ethyl acetate was added thereto, shaken well, and then thoroughly separated into two layers. The upper layer (ethyl acetate layer) was taken. The lower layer (water layer) is again extracted twice with a separatory funnel. The separated upper layers were combined and concentrated under reduced pressure at 50 ° C. using a distillation apparatus equipped with a cooling condenser and dried to obtain a dry weight of 24.9 g.

(2) Rat bean Preparation of extract

Rhynchosia Nolubilis ) was washed with purified water and dried, and then 200 g of the granulated Seomoktae flour was added to 1 liter of an aqueous 70% ethanol solution, boiled for 12 hours in an extractor equipped with a cooling condenser, extracted with 300 mesh filter cloth, and filtered. ^o Leave for 7 days at C to mature and filter with Whatman No. 2 filter paper. The extract was added to a 3-liter separatory funnel, and 1 liter of ethyl acetate was added thereto, shaken and rotted well, and then completely separated into two layers. The upper layer (ethyl acetate layer) was taken. The lower layer (water layer) is again extracted twice with a separatory funnel. The combined upper layers were combined and concentrated under reduced pressure at 50 ^° C. using a distillation apparatus equipped with a cooling condenser and dried to obtain a dry weight of 12.7 g.

Experimental group Sample Preparation

Cell culture Keratinocyte cells (Keratinocyte cells; Kc (Invitrogen; Cascade Biologics)) were grown in medium containing HKGS supplement (Invitrogen; cascade Biologics) in Epilife media (Invitrogen; cascade Biologics). ELISA seeded 150000 Kc cells in a collagen IV coated 12 well plate and seeded 30000 cells on an IV coated 8 well chamber slide. 1uM of retinoic acid, 0.1,1,5% pine needle extract, 0.1, 1, 10% rodent bean extract, 0.1,1,5% mixed extract of pine needles and rodent bean with 0.1,1,5% dissolved in DMSO 1000x stock Made with. The next day, the samples prepared in Kc were diluted to 1000 ×, treated with cells and incubated at 37 ° C. for two days. After 2 days, each plate and chamber slides were washed twice with PBS, irradiated with UVB 13mJ / cm ² , and then added Epilife medium again and sampled after further incubation at 37 ° C. for 4 hours.

Histidase Gene expression assessment ( qPCR data) : Untreated group ( cont ), RA Treatment Group and Efficacy Material Treatment Group

Gene expression levels were measured to determine whether each rat bean extract, pine needle extract and a mixture of both maintain histidase expression when compared with RA (Ratinoic Acid).

After keratinocyte cells were grown using epilife medium, 400000 cells were seeded in 6 well plates. The next day, the samples prepared in Example 2 were diluted 1000-fold each to treat the material and incubated for 2 days, after which the cells were harvested. 500 μl of Trizol was added to each cell, and the cells were homogenized at room temperature for 5 minutes. Add 100ul of chloroform to each tube, shake for 15 seconds by hand, leave for 3 minutes at room temperature, and centrifuge for 15 minutes at 4 ℃, 15000 xg. Only the supernatant was transferred to a new tube, and then 250 μl of isopropyl alcohol was added and mixed well. The mixture was left at room temperature for 10 minutes, and then centrifuged at 4 ° C. and 15000 xg for 10 minutes. After leaving only the pellets (pellets), the supernatant was discarded, and 500 ul of 75% alcohol was added, followed by vortexing. Thereafter, the mixture was centrifuged at 7500 xg for 5 minutes at 4 ° C., and the supernatant was carefully discarded, leaving only pellets. After drying well, deionized water was added to dissolve RNA and quantitated at 260 nm. 4 ug of total RNA (total RNA) was added 2ul of oligo dt (oligo dT) and reacted at 70 ° C. for 10 minutes and then cooled quickly. After DTT, dNTP, 10x RT buffer, MgCl ₂ , RNAase Out 2 minutes at 42 ℃, Superscript III RT was added and reacted at 50 ℃ 60 minutes. QPCR was performed using the synthesized cDNA. At this time, a commercial primer (Hs00157887_m1 *) manufactured by Taqman was used.

As a result (Fig. 1), both rats and pine needles extract was confirmed to maintain histidase expression, in particular, the mixture of rats and pine needles extract mixed 1: 1 with hist similar to the control It was observed to maintain the amount of expression of tidase. Therefore, it can be confirmed that the production of t-UCA is maintained by the soybean extract and pine needle extract or a mixture thereof.

CPD ELISA : Untreated group ( cont ), UVB Treatment Group, UVB Treatment + RA group, UVB Treatment + Efficacy Material Treatment Group

When UVB was irradiated with soybean extract, pine needle extract, and a mixture of both, it was compared with RA (Ratinoic Acid) to determine the degree of DNA damage. Experimental methods for each of the experimental groups are as follows, and each experimental group was prepared in Example 2.

Kc sampled in 12 well plates was extracted using a QIAamp Blood kit (QIAGEN, Cat. No. 51104) and quantified at 260 nm. Each DNA was diluted with PBS to 0.2ug / ml, and then DNA was prepared. The DNA was heated for 10 minutes at 100 ° C for DNA denaturation, and then rapidly cooled in ice for 15 minutes. The plates used for ELISA were precoated 96 well plates the day before with 0.003% protamin sulfate solution. Denatured DNA was added to the wells by 50ul (n = 4), and dried completely at 37 ° C. with O / N. The next day, the DNA coated plate was washed with PBS-T (0.05% Tween-20) 5 times, 150ul of 2% FBS was added to each well, and then incubated at 37 ° C for 30 minutes. Plates were washed 5 times with PBS-T (0.05% Tween-20), CPDs (Anti-cyclobutane pyrimidine dimer; Cosmo bio co.LTD, # NMDND001, Clone: TDM-2) diluted 1: 1000 in PBS 100ul was added to each well and incubated for 30 minutes at 37 ℃. Plates were washed 5 times with PBS-T (0.05% Tween-20), and then Biotin-F (ab ') fragments of IgG (H + L) (1: 2000) in PBS. Diluted to 100ul each well and incubated for 30 minutes at 37 ℃. The plate was washed 5 times with PBS-T (0.05% Tween-20), and Peroxidase-streptavidine was diluted 1: 10000 in PBS, put 100ul into each well and incubated at 37 ° C for 30 minutes. Plates were washed five times with PBS-T (0.05% Tween-20) and once more with citrate-phosphate buffer (pH 5.0). 100ul of substrate buffer was added to each well and incubated at 37 ° C for 30 minutes, and 50ul of 2M H ₂ SO ₄ was added to stop the enzyme reaction, and the absorbance was measured at 492nm.

As a result (FIG. 2), the amount of CPDs increased by ultraviolet (UVB). In addition, when processing the RA, it was confirmed that more CPD is generated. MED is the expression of erythema caused by ultraviolet rays in human body, and CPD, which is an indicator of UV damage in vitro, is known to correlate with MED. Based on these findings, the increase in CPD caused by RA was found to cause erythema, a phenomenon of photosensitivity, when retinol was used during the day. When irradiated with UV light to the rat eye extract and pine needle extract, it was confirmed that CPD is produced at a level similar to or smaller than that of UV, and depending on the concentration, the CPD was produced at a control level. It was observed to have. In addition, it was confirmed that especially when the rat eye extract and pine needle extract showed a similar CPD production inhibitory effect as the control.

Anti-aging efficacy evaluation

After culturing 70 aged fibroblasts in medium106 with low serum growth supplement (LSGS) added to medium106, 60 mm dish 500000 cells were seeded. The next day, the samples prepared in Example 2 were diluted 1000-fold each to treat the material and incubated for 8 hours before harvesting cells. 500 μl of Trizol was added to each cell, and the cells were homogenized at room temperature for 5 minutes. Add 100ul of chloroform to each tube, shake for 15 seconds by hand, leave for 3 minutes at room temperature, and centrifuge for 15 minutes at 4 ℃, 15000 x g. Only the supernatant was transferred to a new tube, 250 μl of isopropyl alcohol was added thereto, and then mixed well. The mixture was left at room temperature for 10 minutes, and then centrifuged at 4 ° C. and 15000 x g for 10 minutes. After leaving only the pellets (pellets), the supernatant was discarded, and 500 ul of 75% alcohol was added, followed by vortexing. Thereafter, centrifugation was performed at 4 ° C. and 7500 × g for 5 minutes, and then the supernatant was carefully discarded, leaving only the pellets. After drying well, deionized water was added to dissolve RNA and quantitated at 260 nm. 4 ug of total RNA (total RNA) was added 2ul of oligo dt (oligo dT) and reacted at 70 ° C. for 10 minutes and then cooled quickly. After DTT, dNTP, 10x RT buffer, MgCl 2, RNAase Out 2 minutes at 42 ℃, Superscript III RT was added and reacted at 50 ℃ 60 minutes. QPCR was carried out using the synthesized cDNA, at which time Taqman's commercial primer (TIMP-1 Hs00171558-m1 * MMP-1 Hs00899658_m1) was used.

As a result (Fig. 3), it was confirmed that the expression of MMP-1 is expressed more in the elderly than the newborn, RA was confirmed that inhibits the expression of MMP-1. In other words, rat eye extract and pine needle extract were confirmed to effectively inhibit the expression of MMP-1 in the elderly to the neonatal level. Hereinafter, one example of the formulation of the composition, but not intended to limit the present invention is intended to be described in detail only.

[Formulation Example 1] Nourishing Lotion (Milk Lotion)

The anti-aging composition according to one embodiment of the present invention was formulated with a nourishing lotion (milk lotion) containing the composition of Table 1 below.

Compounding ingredient weight% Pine Needle Extract 5.0 0 5.0 Rat Bean Extract 0 10.0 5.0 Squalane 5.0 5.0 5.0 Wax 4.0 4.0 4.0 Polysorbate 60 1.5 1.5 1.5 Sorbitan sesquioleate 1.5 1.5 1.5 Liquid paraffin 0.5 0.5 0.5 Caprylic / Capric Triglycerides 5.0 5.0 5.0 glycerin 3.0 3.0 3.0 Butylene glycol 3.0 3.0 3.0 Propylene glycol 3.0 3.0 3.0 Carboxyvinyl polymer 0.1 0.1 0.1 Triethanolamine 0.2 0.2 0.2 Preservative, coloring, fragrance Suitable amount Suitable amount Suitable amount Purified water to 100 to 100 to 100

Formulation Example 2 Flexible Lotion (Skin Lotion)

The anti-aging composition according to one embodiment of the present invention was formulated with a flexible lotion (skin lotion) containing the composition of Table 2 below.

Compounding ingredient weight% Pine Needle Extract 5.0 0 5.0 Rat Bean Extract 0 10.0 5.0 glycerin 3.0 3.0 3.0 Butylene glycol 2.0 2.0 2.0 Propylene glycol 2.0 2.0 2.0 Carboxyvinyl polymer 0.1 0.1 0.1 PEG 12 nonyl phenyl ether 0.2 0.2 0.2 Polysorbate 80 0.4 0.4 0.4 ethanol 10.0 10.0 10.0 Triethanolamine 0.1 0.1 0.1 Preservative, coloring, fragrance Suitable amount Suitable amount Suitable amount Purified water to 100 to 100 to 100

[Formulation Example 3] Nourishing cream

The anti-aging composition according to one embodiment of the present invention was formulated with a nutrient cream containing the composition of Table 3 below.

Compounding ingredient weight% Pine Needle Extract 5.0 0 5.0 Rat Bean Extract 0 10.0 5.0 Polysorbate 60 1.5 1.5 1.5 Sorbitan sesquioleate 0.5 0.5 0.5 PEG60 hardened castor oil 2.0 2.0 2.0 Liquid paraffin 10 10 10 Squalane 5.0 5.0 5.0 Caprylic / Capric Triglycerides 5.0 5.0 5.0 glycerin 5.0 5.0 5.0 Butylene glycol 3.0 3.0 3.0 Propylene glycol 3.0 3.0 3.0 Triethanolamine 0.2 0.2 0.2 antiseptic Suitable amount Suitable amount Suitable amount Pigment Suitable amount Suitable amount Suitable amount Spices Suitable amount Suitable amount Suitable amount Purified water to 100 to 100 to 100

[Formulation Example 4] Massage cream

The composition according to one embodiment of the present invention was formulated with a massage cream containing the composition of Table 4 below.

Compounding ingredient weight% Pine Needle Extract 5.0 0 5.0 Rat Bean Extract 5.0 10.0 5.0 Wax 10.0 10.0 10.0 Polysorbate 60 1.5 1.5 1.5 PEG 60 hardened castor oil 2.0 2.0 2.0 Sorbitan sesquioleate 0.8 0.8 0.8 Liquid paraffin 40.0 40.0 40.0 Squalane 5.0 5.0 5.0 Caprylic / capric triglyceride 4.0 4.0 4.0 glycerin 5.0 5.0 5.0 Butylene glycol 3.0 3.0 3.0 Propylene glycol 3.0 3.0 3.0 Triethanolamine 0.2 0.2 0.2 Preservative, coloring, fragrance Suitable amount Suitable amount Suitable amount Purified water to 100 to 100 to 100

[Formulation Example 5] Pack

The composition according to one embodiment of the invention was formulated into a pack containing the composition of Table 5 below.

Compounding ingredient weight% Pine Needle Extract 5.0 0 5.0 Rat Bean Extract 0 10.0 5.0 Polyvinyl alcohol 13.0 13.0 13.0 Sodium carboxymethylcellulose 0.2 0.2 0.2 glycerin 5.0 5.0 5.0 Allantoin 0.1 0.1 0.1 ethanol 6.0 6.0 6.0 PEG 12 nonyl phenyl ether 0.3 0.3 0.3 Polysorbate 60 0.3 0.3 0.3 Preservative, coloring, fragrance Suitable amount Suitable amount Suitable amount Purified water to 100 to 100 to 100

Formulation Example 6 Gel

The composition according to one embodiment of the present invention was formulated with a gel containing the composition of Table 6 below.

Compounding ingredient weight% Pine Needle Extract 5.0 0 5.0 Rat Bean Extract 0 10.0 5.0 Ethylenediamine sodium acetate 0.05 0.05 0.05 glycerin 5.0 5.0 5.0 Carboxyvinyl polymer 0.3 0.3 0.3 ethanol 5.0 5.0 5.0 PEG 60 hardened castor oil 0.5 0.5 0.5 Triethanolamine 0.3 0.3 0.3 Preservative, coloring, fragrance Suitable amount Suitable amount Suitable amount Purified water to 100 to 100 to 100

As described above in detail the specific parts of the present invention, it is apparent to those skilled in the art that such specific description is merely a preferred embodiment, thereby not limiting the scope of the present invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims

Anti-aging composition comprising at least one substance selected from the group consisting of pine needle extract and decane extract.

The anti-aging composition of claim 1, wherein the pine needle extract is present in an amount of 0.001% to 40% based on the total weight of the composition.

The anti-aging composition of claim 1, wherein the mouse eye extract is contained in an amount of 0.001% to 50% based on the total weight of the composition.

The anti-aging composition of claim 1, wherein the weight ratio of the pine needle extract: rat eye extract is 0.1 to 10: 1.

The anti-aging composition according to any one of claims 1 to 4, wherein the composition is for wrinkle improvement.

The anti-aging composition according to any one of claims 1 to 4, wherein the composition has no photosensitive side effects.

The anti-aging composition according to any one of claims 1 to 4, wherein the composition is a cosmetic composition.

According to claim 7, wherein the composition is softening cream, astringent makeup, nutrition cream, nutrition cream, massage cream, eye cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing water, pack, makeup base, foundation Anti-aging composition comprising a body lotion, body cream, body oil, body essence, body cleansing agent, cosmetic liquid, soap, latex, lotion, ointment, gel, cream, patch or spray.