KR102699167B1 - Oat sprout postbiotic composition having skin whitening and wrinkle improvement activity and use thereof - Google Patents

Abstract

The present invention relates to a sprouted oat postbiotics composition having skin whitening and wrinkle improvement activities and a use thereof. Since the composition has excellent skin whitening and wrinkle improvement effects, it can be usefully utilized as a functional cosmetic composition.

Description

{Oat sprout postbiotic composition having skin whitening and wrinkle improvement activity and use thereof}

The present invention relates to a sprouted oat postbiotics composition having skin whitening and wrinkle improvement activities and its use.

The human skin is located on the outermost part of the body, accounts for a large proportion of the appearance, and is constantly exposed to external stimuli. Recently, as the income and intellectual level of consumers have increased, interest in skin beauty and health has increased to a high level across various age groups, and various cosmetics with more specialized and specific functions such as whitening, wrinkle improvement, and UV protection are being developed.

Human skin color is determined by the concentration and distribution of the black pigment melanin in the skin, and melanin is synthesized in melanocytes in the epidermal layer of the skin. In the melanosome, an organelle within the melanocyte, tyrosine, an amino acid, is converted into DOPA and dopaquinone by an enzyme called tyrosinase, which then undergoes a non-enzymatic oxidation reaction to produce melanin. If melanin is overproduced, the skin color darkens and freckles and other signs occur, so by inhibiting the activity of tyrosinase in the skin, the production and accumulation of melanin can be suppressed, resulting in skin whitening.

Skin aging is largely caused by intrinsic aging and extrinsic aging. Intrinsic aging is caused by a decrease in the secretion of various hormones that control metabolism, a decrease in the function of immune cells, and a decrease in cell activity as we age. Extrinsic aging is caused by an increase in free radicals and reactive oxygen species due to external factors such as ultraviolet rays and environmental pollution. Such skin aging can occur when the synthesis and decomposition of extracellular matrices such as collagen and elastin in the body are not properly regulated, or when their synthesis decreases as aging progresses, and when the expression of collagenase, an enzyme that decomposes collagen fibers present in the extracellular matrix, and elastase, an enzyme that decomposes elastin, is promoted, which can cause a decrease in skin elasticity and the formation of wrinkles.

Oats ( Avena sativa ) are rich in protein, essential amino acids, and soluble fiber compared to other grains, and avenanthramide, a polyphenol component, is known to exist only in oats. There are four types of avenanthramides: A, B, C, and D, and recent research has verified the whitening effect of avenanthramide C. It is known that oat sprouts, which are green sprouts that grow when oats sprout, have a higher polyphenol content than oats, and the concentration of avenanthramide is also known to be significantly higher after germination than before germination, but research results on the whitening and wrinkle-relief effects using oat sprouts are minimal.

Meanwhile, lactic acid bacteria are widely present in nature and are microorganisms that can produce lactic acid by anaerobically utilizing carbohydrates, while postbiotics are byproducts or lactic acid bacteria cell components produced by lactic acid bacteria during the fermentation process. Recently, postbiotics have been developed for various commercial purposes such as food, nutritional supplements, health functional foods, and pharmaceuticals, but there has been no research on specific strains and fermentation conditions for fermentation of sprouted oats as cosmetics.

Accordingly, the inventors of the present invention conducted research to manufacture a postbiotics composition exhibiting excellent skin whitening and wrinkle improvement activities by inoculating lactic acid bacteria into sprout extract and culturing it, and then using sprout fermented solution of oat sprout, thereby completing the present invention.

Republic of Korea Patent Registration No. 10-2016027

The purpose of the present invention is to provide a cosmetic composition for whitening and improving wrinkles containing fermented sprouted oats.

One aspect of the present invention provides a whitening and wrinkle-improving cosmetic composition containing fermented sprouted oats.

The sprouted oats used in the present invention are young leaves of oats ( Avena sativa ), and may be harvested 7 to 10 days after germination.

According to one specific example of the present invention, the fermented oat sprout may be a product obtained by fermenting an oat sprout extract.

The extract included in the composition according to the present invention can be obtained by adding sprouted oat powder to an appropriate amount of solvent and extracting at 100 to 140°C for 10 to 20 minutes.

The above sprouted oat powder can be used by drying and grinding sprouted oats harvested 7 to 10 days after germination, or by purchasing commercially available sprouted oat powder.

According to one specific example of the present invention, the sprouted oat extract can be extracted with a solvent selected from the group consisting of water, alcohols having 1 to 4 carbon atoms, and mixtures thereof.

The solvent may preferably be water, and more preferably distilled water.

According to one specific example of the present invention, the fermented oat sprouts may be cultured at 35 to 40°C for 40 to 80 hours.

According to one specific example of the present invention, the fermentation may be performed using any one strain selected from the group consisting of Lactobacillus plantarum, Lactobacillus rhamnosus , Lactobacillus casei , and Lactobacillus gasseri, but is not limited thereto.

According to one specific example of the present invention, the fermented oat sprout may be included in an amount of 0.01 to 30.0 parts by weight per 100 parts by weight of the cosmetic composition.

The cosmetic composition of the present invention can be prepared in a conventional formulation in the art, and may be, for example, but is not limited to, in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, and spray.

Furthermore, the cosmetic composition of the present invention may be incorporated into other types of solid devices, such as, for example, hydrogels, adhesive patches, non-adhesive patches, microelectronic patches or facial masks, using techniques known to those skilled in the art, or may be incorporated into other makeup products, such as makeup foundations, for example, fluid foundations and compact foundations, makeup remover lotions, makeup remover milks, concealers, eye shadows, lipsticks, lip protectors, lip glosses and powders.

The cosmetic composition of the present invention may further comprise adjuvants commonly used in the cosmetic field, such as fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestering and chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients commonly used in cosmetics.

The cosmetic composition of the present invention may contain various bases and/or additives necessary and appropriate for the formulation of the dosage form, and may be manufactured by further including known compounds such as nonionic surfactants, silicone polymers, conditioners, fragrances, preservatives, bactericides, oxidation stabilizers, organic solvents, ionic or nonionic thickeners, softeners, antioxidants, free radical scavengers, opacifiers, stabilizers, emollients, silicones, α-hydroxy acids, antifoaming agents, moisturizers, vitamins, insect repellents, fragrances, preservatives, surfactants, anti-inflammatory agents, substance P antagonists, fillers, polymers, propellants, alkalizing or acidifying agents, or colorants, within a range that does not reduce the effectiveness.

The sprouted oat postbiotics composition, which is fermented using the lactic acid bacteria of the present invention and has skin whitening and wrinkle improvement activities, has excellent skin whitening and wrinkle improvement effects, and thus can be usefully utilized as a functional cosmetic composition.

Figure 1: Lactobacillus plantarum This is a graph showing the tyrosinase activity inhibition rate of a postbiotic composition fermented by culturing KCTC3104 and Lactobacillus plantarum KCTC3108.
Figure 2 is a graph showing the tyrosinase activity inhibition rate of a postbiotic composition fermented by culturing Lactobacillus rhamnosus KCTC3237 and Lactobacillus rhamnosus KCTC5033.
Figure 3 is a graph showing the tyrosinase activity inhibition rate of a postbiotic composition fermented by culturing Lactobacillus casei KCTC3109.
Figure 4 is a graph showing the tyrosinase activity inhibition rate of a postbiotic composition fermented by culturing Lactobacillus gasseri KCTC3143.
Figure 5 is a graph showing the inhibition rate of eltastase activity of a postbiotics composition fermented by culturing Lactobacillus plantarum KCTC3104, Lactobacillus plantarum KCTC3108, and Lactobacillus rhamnosus KCTC3237.
Figure 6 is a graph showing the collagenase activity inhibition rate of a postbiotics composition fermented by culturing Lactobacillus rhamnosus KCTC3237, Lactobacillus rhamnosus KCTC5033, and Lactobacillus casei KCTC3109.
Figure 7 is a graph showing the collagenase activity inhibition rate of a postbiotics composition fermented by culturing Lactobacillus gasseri KCTC3143.

Hereinafter, the present invention will be described in more detail through one or more examples. However, these examples are intended to exemplify the present invention and the scope of the present invention is not limited to these examples.

Examples 1 and 2. Lactobacillus plantarum ( Lactobacillus plantarum ) Manufacturing of a postbiotic composition of sprouted oats fermented with

Preparation of sprouted oat ( Avena sativa ) extract by concentration and Lactobacillus plantarum Postbiotic compositions were prepared by culturing KCTC3104 and Lactobacillus plantarum KCTC3108, respectively.

Specifically, sprouted oat powder that had been germinated for 10 days was added to distilled water at concentrations of 2.5% w/w (a), 5% w/w (b), and 10% w/w (c), and then extracted and sterilized in an autoclave at 121°C for 15 minutes to obtain a sprouted oat hot water extract.

1.0× ¹⁰⁹ CFU/㎖ of Lactobacillus plantarum KCTC3104 and Lactobacillus plantarum KCTC3108, washed in distilled water, were injected and cultured in a shaking incubator at 37℃ and 75 rpm for 48 hours. Glucose monohydrate (7.5% w/w) and ammonium sulfate (3% w/w) were added as nutrients for the strains. After culture, the supernatant was filtered through Whatman Grade No. 2 filter paper and a syringe filter with a pore size of 0.2㎛, to prepare Examples 1 and 2. Distilled water containing glucose monohydrate and ammonium sulfate was used as a negative control.

2.5% w/w 5% w/w 10% w/w Lactobacillus plantarum KCTC3104 Example 1(a) Example 1(b) Example 1(c) Lactobacillus plantarum KCTC3108 Example 2(a) Example 2(b) Example 2(c)

Examples 3 and 4. Lactobacillus rhamnosus ( Lactobacillus rhamnosus ) Manufacturing of a postbiotic composition of sprouted oats fermented with

It was prepared in the same manner as Example 1, except that Lactobacillus rhamnosus KCTC3237 and Lactobacillus rhamnosus KCTC5033 strains were used and cultured for 72 hours. Distilled water containing glucose monohydrate and ammonium sulfate was used as a negative control.

2.5% w/w 5% w/w 10% w/w Lactobacillus rhamnosus KCTC3237 Example 3(a) Example 3(b) Example 3(c) Lactobacillus rhamnosus KCTC5033 Example 4(a) Example 4(b) Example 4(c)

Example 5. Lactobacillus casei ( Lactobacillus casei ) Manufacturing of a postbiotic composition of sprouted oats fermented with

It was manufactured in the same manner as Example 1, except that Lactobacillus casei KCTC3109 was used. Distilled water containing glucose monohydrate and ammonium sulfate was used as a negative control group.

2.5% w/w 5% w/w 10% w/w Lactobacillus caseiKCTC3109 Example 5(a) Example 5(b) Example 5(c)

Examples 6 and 7. Lactobacillus gasseri ( Lactobacillus gasseri ) Manufacturing of a postbiotic composition of sprouted oats fermented with

It was prepared in the same manner as Example 1, except that Lactobacillus gasseri KCTC3143 strain was used and cultured for 48 hours (Example 6) or 72 hours (Example 7). Distilled water containing glucose monohydrate and ammonium sulfate was used as a negative control.

2.5% w/w 5% w/w 10% w/w Lactobacillus gasseriKCTC3143
(48h) Example 6(a) Example 6(b) Example 6(c) Lactobacillus gasseriKCTC3143
(72h) Example 7(a) Example 7(b) Example 7(c)

Comparative Examples 1 to 6. Preparation of non-fermented sprouted oat extract

After 10 days of germination, sprouted oat powder was added to distilled water at concentrations of 2.5% w/w (a), 5% w/w (b), and 10% w/w (c), and then extracted and sterilized in an autoclave at 121°C for 15 minutes to obtain a sprouted oat hot water extract. 1.0× ¹⁰⁹ CFU/㎖ of Lactobacillus plantarum KCTC3104, Lactobacillus plantarum KCTC3108, washed in distilled water, Lactobacillus rhamnosus KCTC3237, Lactobacillus rhamnosus KCTC5033, Lactobacillus casei KCTC3109, and Lactobacillus gasseri KCTC3143 were mixed with sprouted oat extract and used immediately before the experiment (Comparative Examples 1 to 6).

2.5% w/w 5% w/w 10% w/w Lactobacillus plantarum KCTC3104 Comparative Example 1(a) Comparative Example 1(b) Comparative Example 1(c) Lactobacillus plantarum KCTC3108 Comparative Example 2(a) Comparative Example 2(b) Comparative Example 2(c) Lactobacillus rhamnosus KCTC3237 Comparative Example 3(a) Comparative Example 3(b) Comparative Example 3(c) Lactobacillus rhamnosus KCTC5033 Comparative Example 4(a) Comparative Example 4(b) Comparative Example 4(c) Lactobacillus caseiKCTC3109 Comparative Example 5(a) Comparative Example 5(b) Comparative Example 5(c) Lactobacillus gasseriKCTC3143 Comparative Example 6(a) Comparative Example 6(b) Comparative Example 6(c)

Experimental Example 1. Measurement of the excellent tyrosinase activity inhibition ability of the sprout oat postbiotics composition

In order to confirm the skin whitening efficacy of Examples 1 to 7 and Comparative Examples 1 to 6, a Tyrosinase Inhibitory Assay, a major enzyme involved in the biosynthesis of melanin, was performed.

Specifically, 660 ㎕ of 0.1 M phosphate buffer at pH 6.8 was injected into each e-tube, 60 ㎕ of Examples 1 to 7 and Comparative Examples 1 to 6 were treated into each e-tube, and 90 ㎕ of tyrosinase (750 U/㎖) was treated into each e-tube. Thereafter, 90 ㎕ of 1.5 mM L-tyrosine was treated into each e-tube and reacted at 37°C for 12 minutes. As a positive control group, 60 ㎕ of kojic acid or arbutin, a whitening raw material, was prepared by treating it in the same manner instead of Examples 1 to 7 and Comparative Examples 1 to 6, and as a negative control group, 60 ㎕ of 0.1 M phosphate buffer at pH 6.8 was additionally injected instead of Examples 1 to 7 and Comparative Examples 1 to 6 and treated in the same manner.

After the reaction was completed, 150 μl was dispensed into each 96-well plate, and the absorbance of dopachrome, an intermediate produced by tyrosinase in the melanin biosynthesis pathway, was measured at 490 nm to determine the tyrosinase activity inhibition rate according to the following formula.

Tyrosinase activity inhibition rate (%) =

a: Absorbance after reaction of the blank sample solution

b: Absorbance after reaction of the sample

a', b': Absorbance measured by replacing tyrosinase with buffer

As a result of the analysis, Examples 1 to 5 showed higher tyrosinase activity inhibition rates than the comparative examples at the same concentration at all concentrations, and showed higher tyrosinase activity inhibition rates than the positive control group, nuruk-san at all concentrations. In addition, Examples 6 to 7 showed higher tyrosinase activity inhibition rates than the comparative example 6 at the same concentration at all concentrations, and showed higher tyrosinase activity inhibition rates than the positive control group, arbutin at all concentrations.

In particular, Example 1(c) showed a 1.75-fold increase compared to Comparative Example 1(c), Example 2(c) showed a 1.84-fold increase compared to Comparative Example 2(c) (Fig. 1), Example 3(c) showed a 3.02-fold increase, Example 4(c) showed a 2.86-fold increase in inhibition rate compared to Comparative Example 4(c) (Fig. 2), and Example 5(b) showed a 1.64-fold increase in inhibition rate compared to Comparative Example 5(b) (Fig. 3). In addition, Example 6(c) showed a 2.86-fold increase in inhibition rate compared to Comparative Example 6(c) and a 1.99-fold increase in inhibition rate compared to arbutin, and Example 7(c) showed a 2.58-fold increase in inhibition rate compared to Comparative Example 6(c) and a 1.92-fold increase in inhibition rate compared to arbutin (Fig. 4).

Through these results, it was confirmed that the skin whitening effect of the oat sprout postbiotics composition was excellent, and among the oat sprout postbiotics compositions fermented with various lactic acid bacteria, the skin whitening effect of the oat sprout postbiotics composition fermented for 72 hours by injecting Lactobacillus rhamnosus KCTC5033 was particularly excellent.

Experimental Example 2. Measurement of the excellent elastase activity inhibition ability of the sprouted oat postbiotics composition

In order to confirm the wrinkle improvement efficacy of Examples 1 to 3 and Comparative Examples 1 to 3, an elastase inhibition assay, which is a major cause of decreased skin elasticity and wrinkle formation due to ultraviolet rays, was performed.

Specifically, 200 ㎕ of 1.015 mM N-succinyl Ala-Ala-Ala-p-nitroanilide dissolved in 0.1232 M Tris-HCl (2-Amino-2-(hydroxymethyl)propane-1,3-diol) at pH 8 was treated in a 96-well plate. 10 ㎕ of Examples 1 to 3 and Comparative Examples 1 to 3 were treated to each well, and then left to stand at 25°C for 10 minutes. Thereafter, 10 ㎕ of elastase (3 U/㎖) was treated to each well, and the reaction was performed at 25°C for 10 minutes. As a positive control group, 10 ㎕ of EGCG (Epigallocatechin-3-gallate), known to have excellent antioxidant activity, was prepared by treating it in the same manner instead of Examples 1 to 3 and Comparative Examples 1 to 3.

After the reaction was completed, the absorbance (B) of para-nitroaniline (4-nitroaniline) was measured at 410 nm, and the elastase activity inhibition rate was measured according to the following formula. The buffer solution used was 0.1232 M Tris-HCl at pH 8, the same as in Examples 1 to 3 and Comparative Examples 1 to 3.

Elastase activity inhibition rate (%) =

A: Absorbance after reaction by adding elastase instead of a sample and adding a buffer solution

B: Absorbance of the sample after reaction with elastase added

C: Absorbance of the sample after reaction by adding buffer instead of elastase

D: Absorbance after reaction with buffer added instead of sample and elastase

As a result of the analysis, Examples 1 to 3 showed higher elastase activity inhibition rates than the comparative examples at the same concentration at all concentrations, and it was confirmed that Examples 2(c) and Examples 3(a) to 3(c) showed higher inhibition rates than the positive control group, EGCG (Fig. 5). In particular, Example 3(c) showed the highest elastase activity inhibition rate among Examples 1 to 3, and increased by 2 times compared to Comparative Example 3(c).

Through these results, it was confirmed that the wrinkle improvement efficacy of the oat sprout postbiotics composition was excellent, and among the oat sprout postbiotics compositions fermented with various lactic acid bacteria, the wrinkle improvement efficacy of the oat sprout postbiotics composition fermented for 72 hours by injecting Lactobacillus rhamnosus KCTC3237 was particularly excellent.

Experimental Example 3. Measurement of excellent collagenase activity inhibition ability of sprouted oat postbiotics composition

In order to confirm the wrinkle improvement efficacy of Examples 3 to 7 and Comparative Examples 3 to 6, a collagenase inhibition assay was performed to decompose collagen fibers present in the extracellular matrix and to determine the main cause of wrinkle formation.

Specifically, 800 ㎕ of 0.1 M Tris-HCl at pH 7 was injected into each e-tube, and 1 ㎎ of azocoll (azo-dye impregnated collagen) was added and dissolved. 100 ㎕ of Examples 3 to 7 and Comparative Examples 3 to 6 were treated into each e-tube, and then 100 ㎕ of collagenase (0.2 ㎎/㎖) was treated and stirred at 37℃ for 1 hour to react. As a positive control group, 100 ㎕ of EGCG was prepared by treating it in the same manner instead of Examples 3 to 7 and Comparative Examples 3 to 6.

After the reaction was completed, each e-tube was centrifuged at 10,000 rpm for 10 minutes, and 200 ㎕ of the supernatant was dispensed into a 96-well plate, and the absorbance was measured at 550 nm. The supernatant contained decomposed collagen, and the collagenase activity inhibition rate was measured according to the following formula. The buffer used was 0.1 M Tris-HCl at pH 7, the same as in Examples 1 to 3 and Comparative Examples 1 to 3.

Collagenase activity inhibition rate (%) =

A: Absorbance after reaction by adding collagenase and replacing the sample with buffer

B: Absorbance of the sample after reaction with collagenase added

C: Absorbance of the sample after reaction with buffer added instead of collagenase

D: Absorbance after reaction with buffer added instead of sample and collagenase

As a result of the analysis, Examples 3 to 7 showed higher collagenase activity inhibition rates than the comparative examples at the same concentration at all concentrations, and higher collagenase activity inhibition rates than the positive control group, EGCG, at all concentrations (Figs. 6 and 7). In particular, Example 6(c) showed the highest collagenase activity inhibition rate of about 69% among Examples 3 to 7.

Through these results, it was confirmed that the oat sprout postbiotics composition has excellent skin wrinkle improvement efficacy, and among the oat sprout postbiotics compositions fermented with various lactic acid bacteria, the oat sprout postbiotics composition fermented for 48 hours by injecting Lactobacillus gasseri KCTC3143 has particularly excellent wrinkle improvement efficacy.

The present invention has been described with reference to embodiments thereof. Those skilled in the art will appreciate that the present invention may be implemented in modified forms without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative rather than a restrictive perspective. The scope of the present invention is set forth in the claims, not in the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (7)

A cosmetic composition for whitening and wrinkle improvement containing a fermented extract of sprouted oats ( Avena sativa ),
A cosmetic composition for whitening and wrinkle improvement, wherein the fermentation is performed using any one strain selected from the group consisting of Lactobacillus plantarum , Lactobacillus rhamnosus, Lactobacillus casei, and Lactobacillus gasseri.
delete In the first paragraph,
A cosmetic composition for whitening and wrinkle improvement, wherein the sprouted oat extract is extracted with a solvent selected from the group consisting of water, alcohols having 1 to 4 carbon atoms, and mixtures thereof.
In the first paragraph,
A cosmetic composition for whitening and wrinkle improvement, wherein the fermented product of the above sprout extract is cultured at 35 to 40°C for 40 to 80 hours.
delete In the first paragraph,
A cosmetic composition for whitening and wrinkle improvement, wherein the fermented product of the above sprout extract is contained in an amount of 0.01 to 30.0 parts by weight per 100 parts by weight of the cosmetic composition.
In the first paragraph,
The above whitening and wrinkle-improving cosmetic composition is a whitening and wrinkle-improving cosmetic composition in any one formulation selected from the group consisting of a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, an emulsion foundation, a wax foundation, and a spray.