KR102332943B1 - A cosmetic composition for improving skin aging, containing an extract derived from Sargassum yezoense as an active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving skin aging containing, as an active ingredient, an extract derived from Japanese algae, which is a kind of seaweed, and more specifically, to an extract derived from Japanese mother-of-pearl, characterized in that it contains an extract derived from Japanese algae as an active ingredient. Disclosed is a cosmetic composition for improving skin aging containing as an ingredient.

Description

A cosmetic composition for improving skin aging, containing an extract derived from Sargassum yezoense as an active ingredient}

The present invention relates to a cosmetic composition for improving skin aging, comprising an extract derived from Japanese algae, which is a kind of seaweed, as an active ingredient.

Skin aging can be divided into intrinsic aging due to time-dependent natural aging and extrinsic aging due to external factors. Among extrinsic aging caused by wind, temperature, pollution, and ultraviolet rays, the aging caused by ultraviolet rays is called photoaging. Continuous UV exposure induces an increase in reactive oxygen species in the skin, leading to photoaging. Ultraviolet rays reaching the skin excessively generate active oxygen in the epidermis, thereby observing pigmentation due to irregular melanogenesis in melanocytes. In addition, the arrangement of keratinocytes is irregularly changed, thereby reducing skin moisture. Keratinocytes stimulated by reactive oxygen species secrete inflammatory cytokines to stimulate fibroblasts in the dermis, thereby promoting DNA binding of the NF-κB pathway and activator protein (AP)-1. The expression of metalloprotease (MMPs) is increased by promoting the DNA binding of AP-1. On the other hand, by reducing the synthesis of type 1 procollagen, as a result, it inhibits collagen biosynthesis and promotes decomposition, thereby forming skin wrinkles and reducing elasticity. Based on these biochemical mechanisms, active oxygen induced by UV irradiation is considered to be the most important cause in photoaging skin.

Therefore, in preventing skin aging and improving wrinkles, it is important to discover an antioxidant that removes active oxygen or a material that inhibits collagen decomposition and promotes collagen synthesis.

1. Republic of Korea Patent Publication No. 10-2025634 2. Republic of Korea Patent Publication No. 10-2020-0038755

In the present invention, the wrinkle improvement effect of the Wamojaban-derived extract is confirmed, and the existence of an active material showing good efficacy in improving skin aging showing the effect is confirmed and identified, and the skin containing the Wamojaban-derived extract as an active ingredient It is an object to provide a cosmetic composition for improving aging.

The cosmetic composition for improving skin aging containing the extract derived from jabanum of the present invention as an active ingredient, which has solved the above problems, 1) prepare a crude extract prepared by first extracting dried jabanum with methanol (MeOH) and drying it step and

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2) 1 kg of the crude extract is extracted three times at room temperature using 3 L of acetone and dried under reduced pressure to obtain the extract. A cosmetic composition for improving skin aging comprising an extract derived from jabanum as an active ingredient, characterized in that
[Formula 2]

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As a result of measuring the activity of the extract and active substances for improving skin aging, the cosmetic composition for improving skin aging, which contains the extract derived from jabanum from the present invention, as an active ingredient, SHQA has a higher degree of activity with respect to PPARα activity. It can be seen that it is very effective in moisturizing the skin by increasing the expression of filaggrin and involucrin.

In addition, it can be seen that by inhibiting the activities of MMP-2 and MMP-9 that affect the epidermal-dermis boundary, it can protect the epidermal-dermis boundary, and it also has the effect of preventing the deterioration of skin elasticity by inhibiting the Elastase activity. could check

In addition, it was found that the expression of MMP-1, which plays a very important role in wrinkle formation by participating in the decomposition of the dermis, was suppressed, and thus the anti-wrinkle effect was also excellent.

In addition, SY extract is effective for multiple targets, that is, it is very effective in improving skin aging, so it is a good material with very good value as a functional cosmetic material for improving wrinkles in the future.

1 is a diagram showing the extraction and fractionation process of jabanum according to an embodiment of the present invention.
FIG. 2 shows chromatograms of the second, third, and fourth subfractions among the subfractions of FIG. 1 .
3 to 5 are HPLC and NMR analysis graphs of compound 1 isolated and identified from the small fractions 2, 3, and 4 of FIG. 2 .
Figure 6 shows the results of PPAR transactivation of jabanum extract according to an embodiment of the present invention.
Figure 7 shows the filaggrin / involucrin expression test results of the genus Capricorn extract according to an embodiment of the present invention.
Figure 8 shows the results of the MMP-2 and MMP-9 activity inhibition efficacy test of the wamojaban extract according to an embodiment of the present invention.
Figure 9 shows the results of the elastase (Elastase) test of the jabanum extract according to an embodiment of the present invention.
Figure 10 shows the results of the MMP-1 and Procollagen expression test of the jasmine extract according to an embodiment of the present invention.
Figure 11 shows the radical scavenging ability (DPPH) test results of the extract according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to preferred experimental examples.

An object of the present invention is to provide a cosmetic composition for improving skin aging, which contains an extract derived from Wamojaban as an active ingredient,

In more detail, the cosmetic composition for improving skin aging containing the extract derived from jabanum of the present invention as an active ingredient comprises the extract from jabanum jabanum as an active ingredient,

The Wamojaban extract is characterized in that it is extracted according to the extraction and fractionation process as shown in FIG. 1, and more specifically, it is extracted by the following method.

1) A step of preparing a crude extract prepared by first extracting the dried jabanon with methanol (MeOH) and drying it.

2) Using 3 L of acetone to 1 kg of the crude extract, extracting three times at room temperature and drying under reduced pressure to obtain an extract.

According to the present invention, the wamojaban extract contains the following Chemical Formulas 1 to 3 as an active material.

[Formula 1]

[Formula 2]

[Formula 3]

According to the present invention, preferably, the wamojaban extract contains the following Chemical Formula 1 as an active material for improving skin aging, wherein the following Chemical Formula 1 contains 10% by weight or more with respect to the total amount of the extract.

[Formula 1]

As a result of the preliminary experiment by the present inventors, the Wamojaban extract according to the present invention confirmed the Wamojaban extract showing good efficacy in improving skin aging from 98 kinds of marine natural product libraries. It promotes the expression of filaggrin and involucrin, which are involved in skin moisturizing, has antioxidant activity, and inhibits the activities of MMP-2 and MMP-9 involved in the decomposition of skin DEJ, and is an elastase that is involved in lowering skin elasticity. It was confirmed that the activity of

In addition, it was confirmed that it inhibits the activity of tyrosinase involved in whitening and also inhibits the activity of α-glucosidase involved in tyrosinase maturation. is judged

In order to confirm the efficacy of the extract for wrinkle improvement constituting the present invention, first, the expression level of MMP-1 and the decomposition level of collagen I were checked, and the index component was checked for setting standards and test methods. , and also performed an efficacy search on the isolated active ingredients to confirm the ingredients showing the activity of Wamojaban.

<Test method>

1. Collection of seaweed and preparation of algae extract

Seaweeds collected near the east coast were identified by comparative analysis through a botanical book, etc., dried, extracted with MeOH, and dried to be used as a crude extract in this study.

2. Isolation and identification of active ingredients

The dried Japanese capsicum (1Kg) was extracted three times at room temperature using 3L of acetone, dried under reduced pressure, the extract (35g) was suspended in water, and fractionated with hexane (Hx) and ethyl acetate (EA) and dried, respectively. 4 g and 30 g of dry fractions were obtained. The double EA fraction was purified by silica gel vacuum solumn chromatography with n-hexane, ethyl acetate and MeOH (10:1:0, 3:1:0, 1:1:0, 0:1:0, 0:10:1). , 0:5:1, and 0:0:1; each 500mL) was divided into 7 subfractions under solvent conditions. Of these, 2, 3, and 4 small fractions (5 g out of 24.8 g) were combined and C ₁₈ reverse phase Prep HPLC [Gilson 321; ^{Phenomenex ® Luna C18 (2) 10} μ mcolumn (21.2 × 250mm); 10mL/min; UV detection at 250nm], 50% - 100% MeCN/H ₂ O (in 0.02% TFA, for 60 min) solvent conditions to extract compound 1 (SHQA 3.5 g), compound 2 (SC), and compound 3 (SQA 40 mg) got it

3. Active search for skin aging

3.1. PPAR Transactivation assay

CV-1 cells, a monkey kidney epithelial cell line, were spread on a 24-hole plate and cultured for 24 hours, followed by transient transfection with three types of plasmid genes: PPAR α/δ/γ, + PPRE-Luc + pRL-SV40. After incubation for 24 hours, candidate substances were treated by concentration, and cells were lysed after incubation for 24 hours to measure the activity of expressed luciferase using a dual-luciferase reporter assay system.

3.2. Measurement of filaggrin/involucrin expression

HaCaT cell line derived from human keratinocyte was aliquoted into 60 mm dishes, cultured for 1 day, treated with each candidate material, and cultured for 24 hours, and total RNA was isolated using Trizol and RT-PCR Filaggrin/involucrin expression levels were compared, and if necessary, protein was isolated, western blot was performed, and density was measured and quantified.

3.3. MMP-2, -9 Activity Inhibition Efficacy Measurement

After treatment with TPA in NHF, MMP-2 or -9 enzyme was induced in the supernatant and used in this experiment. After electrophoresis of the supernatant on the Gelatin zymography gel, it was treated with Buffer1 (2% Triton X-100) for renaturation, and then treated with Buffer2 (50mM Tris pH8.0, 5mM CaCl ₂ , 0.02% NaN ₃ ) and candidate substances simultaneously for 24 hours. After the reaction, the generated band was stained with Coomassie brilliant blue to measure the band density.

3.4. Measurement of Elastase Activity Inhibition Efficacy

After reacting human leukocyte elastase or porcine elastase with a candidate substance and substrate, the amount of decomposed p-nitroaniline was measured for absorbance at 405 nm.

3.5. Effect on MMP-1 and procollagen expression

The amount of MMP-1 or procollagen secreted after stimulation with UVB or TNFα using normal fibroblasts (NHF) or cell line Hs68 cells was measured using ELISA and western blot.

3.6. DPPH free radical scavenging activity

The extract and the sample of the compound were uniformly mixed with a solution diluted in an extraction solvent to an appropriate concentration with a 100 uM DPPH solution, and then left at room temperature for 30 minutes, and then absorbance was measured at 517 nm.

<Test result>

1. Compounds isolated from Dwarf Mojaban

Extraction and fractionation of dwarf hat was carried out with the scheme shown in FIG. 1 .

In the extraction and fractionation, HPLC chromatogram analysis of the 2nd, 3rd, and 4th subfractions of the 7 subfractions of the EA fraction was performed, and the results are as shown in FIG. 2 of the accompanying drawings.

[HPLC Chromatogram Analysis]

HPLC chromatogram using a linear gradient of 10 to 100% aqueous acetonitrile (0.02% TFA) for 30 min ( Column, Phenomenex Luna 15u C18(2) 4.6 x 100 mm; flow, 1ml/min, UV detection at 250nm).

SHQA- t _R 26.423 min SQA- t _R 31.320 min

The separated compound was analyzed for its structure through a Varian 500 MHz spectrometer. As a result of the analysis, compound 1 was identified as sargahydroquinoic acid and compound 3 as sargaquinoic acid. A small peak in the middle was confirmed with sargachromenol. (See attached drawings 3 to 5)

Peak 1

Peek 2

Peak 3

Among the separated compounds, compound 1, sargahydroquinoic acid, accounted for more than 10% of the MeOH extract and was set as an indicator material.

2. Activity test for skin aging

2.1 PPAR Transactivation

First, as a result of measuring the degree of transactivation for PPARα and PPARγ, 75% and 70% of the positive control group were activated when treated with 10ug/ml, respectively.

Effects of SY extracts of PPARα/γ transcriptional activity PPARα (%) PPARγ (%) SY 10ug/ml 75.25 70.35 Wy14,643 100 (4.2 fold) Troglitazone 100 (4.7 fold)

As a result of measuring transactivation for SHQA and SQA, the compounds isolated from SY, SQA showed high activity against PPARγ and SHQA against PPARα. In SY, there is more SHQA to be considered as an index component, so the effect of improving skin aging by SHQA is expected. (See Fig. 6)

2.2. Filaggrin/involucrin expression

As a result of measuring the expression of filaggrin and involucrin, the proteins involved in skin moisturizing, high expression rates were shown in the group treated with SY extract, SHQA, SQA 10 ug/ml, and 10 uM, so it is a feed as a raw material with good skin moisturizing effect. (see Fig. 7)

2.3. MMP-2, -9 activity inhibitory effect

The effect on the inhibition of the activity of MMP-2 and MMP-9, proteins involved in the decomposition of the epidermis and dermis of the skin, was measured by zymography and western blot. The degree of activity and expression inhibition was shown by measuring the density of the band when treated with 100 ug/ml or 100 uM. (See Fig. 8)

2.4. Elastase activity inhibitory effect

For the inhibitory ability of elastase affecting skin elasticity, the inhibitory ability was evaluated at 100 ug/ml or 100 uM using elastase isolated from pigs and humans. It is considered to be a material that can help improve elasticity by showing a much higher inhibitory ability in human elastase. (See FIG. 9)

2.5. Effect on MMP-1 and procollagen expression

The expression of MMP-1, which has a great effect on collagen degradation in the dermal dermis, and the efficacy of inhibiting collagen degradation were measured. In the case of MMP-1, it was confirmed that it was effective in preventing wrinkles by reducing the amount of expression, and in the case of SY and SHQA, the activity was stronger. In the case of collagen, it was confirmed that the effect of preventing decomposition was not very strong. (See Fig. 10)

2.6. Efficacy on radical scavenging ability

The efficacy of SY, SHQA, and SQA on radical scavenging activity was measured by the DPPH method. The inhibitory ability and IC ₅₀ values at 100 ug/ml were obtained. As a result of the measurement, it was confirmed that the radical scavenging ability was very excellent. (See Fig. 11)

From the above test results, the present inventors judged that it is preferable to select sargahydroquinoic acid (SHQA), which accounts for 10% or more of the extract, as the indicator component of the serrano jasmine extract (SY) selected as a skin aging improvement material.

In addition, as a result of measuring the activity of the extracts and compounds for skin aging improvement, it was estimated that SHQA had a higher level of activity for PPARα activity, and it was very effective in moisturizing the skin by increasing the expression of filaggrin and involucrin. could

In addition, it was confirmed that it can protect the epidermal-dermis boundary by inhibiting the activities of MMP-2 and MMP-9 that affect the epidermal-dermis boundary, and it was confirmed that it can also prevent the deterioration of skin elasticity by inhibiting the elastase activity. In addition, it was confirmed that the anti-wrinkle effect was also excellent by suppressing the increase in the expression of MMP-1, which plays a very important role in the generation of wrinkles by participating in the decomposition of the dermis.

The SY extract had an inhibitory effect on tyrosinase and α-glucosidase, so it was predicted to have a whitening effect.

As a result of the above confirmation, it is judged that SY can be used as a good material worthy of development as a functional cosmetic material for wrinkle improvement in the future because it is effective in multiple targets, that is, it is very effective in improving skin aging.

Claims (4)

delete 1) preparing a crude extract prepared by first extracting the dried jabanon with methanol (MeOH) and drying;
2) 1 kg of the crude extract is extracted three times at room temperature using 3 L of acetone and dried under reduced pressure to obtain the extract. A cosmetic composition for improving skin aging comprising an extract derived from jabanum as an active ingredient, characterized in that
[Formula 2]

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