KR102134150B1 - A composition comprising extract of Prasiola japonica for improving skin wrinkle caused by photoaging - Google Patents

Abstract

The present invention relates to a composition for improving wrinkles comprising an extract of fresh water ( Prasiola japonica ) as an active ingredient. The freshwater seaweed ( Prasiola japonica ) extract has the effect of reducing the expression of collagen degrading enzymes MMP-1 and MMP-3 in photo-aging-induced dermal keratinocytes (HaCaT cells) and increasing the expression of Sirt-1, which increases collagen production. Excellent, it can be usefully used as a composition or cosmetic composition for improving skin wrinkles by photoaging.

Description

A composition comprising extract of Prasiola japonica for improving skin wrinkle caused by photoaging}

The present invention relates to a composition for improving wrinkles comprising a rollolide as an active ingredient.

There are two main causes of skin aging. The first is natural aging that progresses over time (intrinsic aging), and the second is extrinsic aging due to skin damage caused by external environment and irritation. Natural aging is influenced by genetic factors, so artificial control is difficult, whereas exogenous aging is influenced by environmental factors, so artificial control is relatively easy. In particular, the aging phenomenon caused by ultraviolet rays as an exogenous aging phenomenon is called a photoaging phenomenon (Lee, J. N. et al., J. Soc. Cosmet. Sci. Korea., 32(4), 227-231, 2006).

Photoaging causes acute and chronic skin injury, such as skin cancer, by continuous exposure to UV light. According to the wavelength range existing in sunlight, UV is divided into three types: UV-A (320-400 nm), UV-B (290-320 nm) and UV-C (200-290 nm), of which UV-C Is almost completely absorbed and filtered by the ozone layer, whereas UV-A and UV-B reach the earth's surface, so UV-A and UV-B are known to be the main factors inducing photoaging (Pandel, R. et al., ISRN Dermatol., 2013, 930164, 2013). UV-B has a higher mutation rate and contains more energy than UV-A, which results in more detrimental effects (Yaar, M. et al., Br J Dermatol., 157(5), 874-887, 2007). In addition, UV-B may cause various side effects such as decreased collagen and vascular dilation of the skin due to an increase in the thickness of the epidermis, the elasticity of the skin, and matrix metalloproteinases (MMPs) after passing through the epidermis of the skin. . As a result, precancerous lesions such as erythema, cell overproduction, wrinkle formation and photoaging appear to be important causes of skin cancer.

One of the most prominent features of skin aging is wrinkle formation, which is caused by reduction or inhibition of collagen present in the dermal layer. In particular, inhibiting the production of collagen is important for maintaining the structure and shape of the dermis, and means that the level of collagen fibers that provide firm strength to the skin cannot be maintained normally, and as a result, collagen is reduced to form wrinkles on the outside of the skin. Will appear.

The mechanism of wrinkle formation is not fully understood, but there is a difference in the process of wrinkle formation by natural aging and photoaging. Wrinkle formation due to natural aging is caused by a decrease in physiological function. The action of fibroblasts and the number of cells in the dermal tissue decreases, reducing the amount of synthesis of extracellular matrix (ECM) such as collagen and elastin. Becomes loose. As a result, elasticity is reduced, moisture in skin cells is lost, and the structure of the stratum corneum changes, resulting in fine lines.

On the other hand, skin wrinkle formation due to photoaging accelerates skin aging by preventing skin fibroblasts from producing collagen by ultraviolet rays. In addition, ultraviolet rays increase the expression of matrix metalloproteinases (MMPs), which are collagen degrading enzymes, to promote the decomposition of collagen fibers and other fibers that make up the skin. These phenomena appear more in skin exposed to UV rays than natural aging skin, and cause many deformations and damages to the structure and shape of the dermis, and eventually more severe wrinkles are formed on the skin (Jin Soo Nam, J. Soc et al. Cosmet. Sci. Korea., 40(4), 321-330, 2014).

MMPs are secreted from many cells, including keratinocytes and fibroblasts, and maintain skin elasticity by decomposing most of the protein components that make up the extracellular matrix and basement membrane (BM). It is known to destroy connective tissue and cause wrinkles, deterioration of elasticity, and sagging of the skin. Recently, the cell signaling pathway related to the increase of MMPs with aging has been reported to be mainly involved in the MAP kinase pathway, and the most affected factor in this pathway is the AP-1 (activator protein-1) factor, which is the growth and differentiation of cells. It regulates the expression of many genes involved in and strongly regulates the expression of some MMPs (Hyeji Jeon et al., Korean J. Microbiol. Biotechnol., 41(4), 449-455, 2013).

Among the MMPs whose expression is regulated by AP-1, matrix metalloproteinase-1 (MMP-1) is a protease present in the skin, and is an important protein for cell-to-cell morphogenesis, wound healing, tissue regeneration, and reorganization. However, when exposed to ultraviolet rays, MMP-1 is produced in the body more than usual, and after binding with collagen, collagen is cut to cause skin aging. Therefore, it can be said that skin exposure by UV rays activates the MAP kinase pathway to induce the expression of AP-1 and increase the expression of MMPs to decompose the extracellular matrix to promote wrinkle formation (So Seung-ho et al., J. Ginseng Res ., 32(1), 39-47, 2008).

Sirtuin is a protein deacetylase made in some tissues of the body, such as the brain, liver, and kidneys. In particular, for Sirt-1, diseases such as inflammation, apoptosis, diabetes and aging Is importantly involved. In addition, prior literature [Han, J. G., Ph.D. dissertation, Yonsei University, 2015; Kim, J. et al., Mol Cell Endocrinol., 412, 216-225, 2015; Serravallo, M. et al., Arch Dermatol Res., 305(4), 269-282, 2013] showed that activation of Sirt-1 helps collagen production and can be used as an anti-aging material effective in wrinkle improvement. It started.

Currently, retinoic acid and retinol have been proven to inhibit skin wrinkle formation, and dehydroepiandrosterone, ginsenoside Rg3, and flavonoids have been reported to improve wrinkles and protect skin. In the skin aging phenomenon, even if natural aging is inevitable, it is a method to delay and suppress skin aging by effectively blocking photoaging caused by ultraviolet rays, which is an environmental factor.

Unlike fresh seaweed, Prasiola japonica is a green algae (Phylum Chlorophyta). It grows in small rivers, not in the sea, and belongs to the Freshwater Parasiolaceae family (Park, MK et al., J. Plant Bio., 13, 1-10, 1970). It has been reported that freshwater seaweed contains mannose, loliolide, sorbitol, glucitol, mannitol and alveine that have not been observed in seaweed. (Seo, DW et al., J. Biomed. Res., 14(2), 83-90, 2013).

Loliolide inhibits the activity of acetylcholinesterase (Fang, Z. et al., Chem Pharm Bull (Tokyo)., 58(9), 1236-1239, 2010), and human colorectal cancer cells Anti-proliferative activity in host Caco-2 (Machado, FB et al., Molecules, 17(2), 1852-1859, 2012) has been reported.

As a prior document related to a composition for improving wrinkles containing lololilide, Korean Patent No. 10-1437729 discloses a composition for inhibiting cell aging containing lololilide as an active ingredient, and prior literature [Yang, X. et al., The Korean Society of Phycology, 26(2), 201-208, 2011.

In the case of the Korean Patent No. 10-1437729, senescence-associated β-galactosidase (SA-β), which is a representative indicator of cell aging in natural senescent cells (aged fibroblasts induced by adriamycin) -gal) The cell aging inhibitory effect associated with cell life was confirmed from the activity inhibitory effect, and the prior literature [Yang, X. et al., The Korean Society of Phycology, 26(2), 201-208, 2011] In this case, the effect of protecting cell damage or cell death was confirmed from antioxidant activity in VERO cells (Monkey kidney fibroblast cells), not skin cells.

In the present invention, the effect of improving wrinkles was confirmed from the changes in the expression levels of MMP-1, MMP-3 and Sirt-1 according to the treatment of lololilide in the skin keratinocytes (HaCaT cells) induced by photoaging (UV-B), As a result, it was found that lololilide is a composition that has a much more selective effect on improving wrinkles due to photoaging than surprisingly improving wrinkles caused by natural skin aging. However, there has been no previous report confirming that the rollolide, as in the present invention, is a composition that exhibits a selective wrinkle improvement effect on photoaging.

Korean Registered Patent No. 10-1437729, a composition for inhibiting cell aging containing lorioride as an active ingredient, registered on August 28, 2014.

Nam Jin-Joo et al., Efficacy of skin extracts and their fractions from the extract of Nungsil, J. Soc. Cosmet. Sci. Korea., 40(4), 321-330, 2014. So Seung-ho et al., Mechanism of Action on Skin Wrinkle Improvement of Red Ginseng Herbal Medicine Complex (KTNG0345), J. Ginseng Res., 32(1), 39-47, 2008. Wrinkle improvement effect of Juniper tree (Juniperus rigida Sieb.), Korean J. Microbiol. Biotechnol., 41(4), 449-455, 2013. Fang, Z. et al., Anticholinesterase and antioxidant constituents from Gloiopeltis furcata, Chem Pharm Bull(Tokyo)., 58(9), 1236-1239, 2010. Han, J. G., Anti-aging Effects of a Novel Tripeptide Derived from Chlorella vulgaris in Human Skin Cells: Effect of New Tripeptide from Chlorella vulgaris on Anti-aging of Human Skin Cells, Doctoral Thesis, Yonsei University, 2015. Kim, J. et al., The natural phytochemical dehydroabietic acid is an anti-aging reagent that mediates the direct activation of SIRT1, Mol Cell Endocrinol., 412, 216-225, 2015. Lee, J. N. et al., Anti-wrinkle effect of Morinda citrifolia (Noni) extracts, J. Soc. Cosmet. Sci. Korea., 32(4), 227-231, 2006. Machado, F. B. et al., Evaluation of the antiproliferative activity of the leaves from Arctium lappa by a bioassay-guided fractionation, Molecules, 17(2), 1852-1859, 2012. Pandel, R. et al., Skin photoaging and the role of antioxidants in its prevention, ISRN Dermatol., 2013, 930164, 2013. Park, M. K. et al., Ecological and morphological studies on the Prasiola sp. in the Samchuck-Chodang, J. Plant Bio., 13, 1-10, 1970. Seo, D. W. et al., Screening of functional components derived from fresh water laver, Prasiola japonica, and its pharmacological properties, J. Biomed. Res., 14(2), 83-90, 2013. Serravallo, M. et al., Sirtuins in dermatology: applications for future research and therapeutics, Arch Dermatol Res., 305(4), 269-282, 2013. Yaar, M. et al., Photoageing: mechanism, prevention and therapy, Br J Dermatol., 157(5), 874-887, 2007. Yang, X. et al., Antioxidant activity and cell protective effect of loliolide isolated from Sargassum ringgoldianum subsp. coreanum, The Korean Society of Phycology, 26(2), 201-208, 2011.

An object of the present invention is to provide a composition for improving wrinkles, which comprises a rollolide as an active ingredient.

The present invention relates to a composition for improving wrinkles or a cosmetic composition for improving wrinkles due to photoaging, which includes lololilide as an active ingredient.

The lololilide may be synthesized according to a conventional method, or may be separated and purified from freshwater seaweed ( Prasiola japonica ). The freshwater seaweed is a green algae that is different from seaweed, which is a red algae, and is not observed in seaweed, mannose, loliolide, sorbitol, glucitol, mannitol, and al. Contains alveine.

The cosmetic composition of the present invention is preferably from 0.001% by weight to 50% by weight, more preferably from 0.001% by weight to 40% by weight, most preferably from 0.001% by weight to 30% by weight relative to the total weight of the total composition of the cosmetic composition. %.

The cosmetic composition may be prepared in any formulation conventionally prepared in the art, for example, solution, emulsion, suspension, paste, cream, lotion, gel, powder, spray, surfactant-containing cleansing, oil , Soap, liquid detergent, bath, foundation, makeup base, essence, lotion, foam, pack, soft water, sunscreen cream and sun oil may be prepared in a formulation selected from the group consisting of, but is not particularly limited thereto.

In addition, the cosmetic composition may include all of the components generally used in cosmetics, in addition to lololiide. For example, it may include general auxiliary ingredients such as emulsifiers, thickeners, emulsions, surfactants, lubricants, alcohols, water-soluble polymers, gelling agents, stabilizers, vitamins, inorganic salts, emulsifiers, and fragrances. The above ingredients can be selected depending on the formulation or purpose of use, within the range that does not impair the inherent effects of cosmetics.

The cosmetically acceptable carrier included in the cosmetic composition of the present invention may vary depending on the formulation of the cosmetic composition.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol , 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan and the like can be used.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, suspensions such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, micro Crystalline cellulose, aluminum metahydroxide, bentonite, tracant, or the like can be used.

When the formulation of the present invention is a paste, cream, or gel, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. are used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used as a carrier component, and in the case of a spray, additionally, chlorofluorohard Propellants such as chlorocarbon, propane/butane or dimethyl ether.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide as a carrier component Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

The cosmetic composition containing the rollolilide can be used daily and can be used for an indefinite period, preferably, according to the user's age, skin condition or skin type, the concentration of the rollolilide, the number of uses, and You can adjust the duration.

The present invention relates to a composition for improving wrinkles comprising a rollolide as an active ingredient. The lololilide has excellent expression reduction effect of collagen degrading enzymes MMP-1 and MMP-3 and photoinitiation of Sirt-1, which increases collagen production, in photo keratinocytes (HaCaT cells) where photoaging is induced. It can be usefully used as a composition or cosmetic composition for improving skin wrinkles.

1 is a result of confirming the cytotoxicity according to the treatment with lololilide in HaCaT cells.
Figure 2 is a result of confirming the mRNA expression level of MMP-1 according to freshwater seaweed extract (FIG. 2A) or lololiide (FIG. 2B) treatment in aging-induced HaCaT cells.
Figure 3 is a result of confirming the mRNA expression level of MMP-3 according to freshwater seaweed extract (FIG. 3A) or lololiide (FIG. 3B) treatment in aging-induced HaCaT cells.
Figure 4 is a result confirming the mRNA expression level of Sirt-1 according to the treatment with lololilide in HaCaT cells induced photoaging.

Hereinafter, a preferred embodiment of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the contents introduced herein are thorough and complete, and are provided to sufficiently convey the spirit of the present invention to those skilled in the art.

<Example 1. Culture of HaCaT cells>

Human keratinocyte cell (HaCaT) cells were cultured at 37° C. using DMEM (Dulbecco's Modified Medium) medium containing 10% fetal bovine serum (FBS), 1% penicillin, and 100 U/ml streptomycin. It was used for the experiment by subculturing in 5% CO2 condition.

<Example 2. Confirmation of cytotoxicity of lololilide>

The cytotoxicity according to the treatment of the lololilide of the present invention was confirmed by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay method.

The HaCaT cells passaged in Example 1 were dispensed at 100 µl in a 96-well plate to be 1×10 ⁵ cells per well, and each well had 0, 25 lololilide (chemfaces, Wuhan, Hubei, China). , 50 and 100 μM and incubated for 24 hours. After 24 hours, 100 µl of the supernatant of the culture was removed, and 10 µl of a 5 mg/ml MTT solution was added, and the mixture was reacted at 37° C. for 3 hours. Thereafter, the reaction solution was removed, and 100 µl of the reaction stop solution (10% SDS in 0.1N HCl) per well was reacted for 24 hours, and absorbance was measured at 540 nm, and the results are shown in FIG. 1. At this time, based on the cell viability of the untreated HaCaT cells based on 100%, the viability of the cells treated with the sample was calculated.

Referring to FIG. 1, when it was treated with HaCaT cells, it was found that irrespective of the concentration, the cell viability was similar to that of the control group (untreated group), indicating no cytotoxicity.

<Example 3. Confirmation of wrinkle improvement effect of lololilide>

In order to confirm the wrinkle improvement effect by photoaging, mRNA expression levels of MMP-1, MMP-3, and Sirt-1 according to treatment of UV-B and lololilide were confirmed.

HaCaT cells passaged in Example 1 above were dispensed in 2 ml so as to be 8×10 ⁵ cells per well in a 6-well plate, and cultured for 24 hours. After 24 hours, the culture was removed, and then 1 ml of PBS was treated, and photoaging was induced by irradiating 30 mJ/cm 2 of UV-B with a UV-B lamp. After UV-B irradiation, 10% FBS, 1% penicillin, and 1 ml of DMEM medium to which 100 U/ml streptomycin was added, and the extract of rololilide of Example 1 or a control freshwater ethanol aqueous solution were treated by concentration.

Subsequently, mRNA expression levels of MMP-1 (FIG. 2), MMP-3 (FIG. 3), and Sirt-1 (FIG. 4) according to lololiide treatment in photo-induced HaCaT cells were confirmed, FIGS. 2 to 4 In the comparison group, H ₂ O ₂ was treated with 50 μM instead of UV-B irradiation to induce aging, and thus shown in FIGS. 2 and 3.

2 to 4, in HaCaT cells in which photoaging is induced by UV-B, the present invention, lololilide, is a collagen degrading enzyme MMP-1 (FIG. 2) and MMP-3 ( The mRNA expression level of FIG. 3) is excellent, but the mRNA expression level of Sirt-1 (FIG. 4) related to collagen biosynthesis increases in a concentration-dependent manner, and it can be seen that lololilide is an effective composition for improving wrinkles due to photoaging.

Particularly, compared to aging caused by H ₂ O ₂ , lololiide has a better effect of reducing mRNA expression levels of MMP-1 and MMP-3 in a group in which photoaging is induced, and thus, lololilide is effective in improving wrinkles caused by photoaging. It was found to be a more selective composition.

<Formulation Example 1. Preparation of cosmetic composition>

Formulation Example 1-1. Preparation of flexible lotion

Using the lololiide of the present invention, according to the composition shown in Table 1, a flexible lotion was prepared by a conventional method.

Raw material name Content (% by weight) Lololilide 4.0 Butylene glycol 3.5 glycerin 2.5 Polyoxyethylene cured castor oil 0.1 ethanol 2.5 Betaine 1.0 Citric acid 0.01 Sodium citrate 0.03 antiseptic Proper Spices Proper Purified water to 100

Formulation Example 1-2. Preparation of nutritional essence

Nutritional essences were prepared by the conventional method according to the composition shown in Table 2 below using the lololilide of the present invention.

Raw material name Content (% by weight) Lololilide 7.0 Cetostearyl alcohol 1.0 Self emulsifying monostearic acid 1.0 Wax 0.5 Squalane 5.0 Isocetyl octanoate 3.0 Dimethylsiloxane 0.3 Sorbitan monostearate 0.5 Polyethylene glycol monostearate 8.0 glycerin 4.0 Propylene glycol 0.2 Carboxy polymer 0.22 Triethanolamine 0.25 antiseptic Proper Spices Proper flash Proper Purified water to 100

Formulation Example 1-3. Manufacture of cream

The cream was prepared by a conventional method according to the composition shown in Table 3 below using the lololilide of the present invention.

Raw material name Weight% (w/w) Lololilide 7.0 Cetostearyl alcohol 3.0 Self-emulsifying monostearic acid 1.5 Lipophilic monostearic acid 1.5 Wax 0.5 Floating paraffin 8.0 Squalane 7.0 Isocetyl octanoate 4.0 Refined jojoba oil 4.0 Dimethylsiloxane 0.3 Sorbitan monostearate 1.0 Polyethylene glycol monostearate 1.2 glycerin 6.0 Propylene glycol 4.0 Betaine 4.0 Shotgun Sword 0.06 Triethanolamine 0.10 antiseptic 0.25 Spices Proper flash Proper Purified water to 100

Claims (3)

In the cosmetic composition for improving wrinkles by photoaging, comprising a freshwater kim (Prasiola japonica) extract as an active ingredient, wherein the freshwater kimchi extract is an ethanol extract, the cosmetic composition for improving wrinkles by photoaging. delete delete

Images