KR102816096B1 - Food composition and cosmetic composition for skin anti-aging and moisturizing - Google Patents

Abstract

The present invention relates to a food composition and a cosmetic composition for skin anti-aging and moisturizing, comprising a glycine-phenylalanine derivative as an effective ingredient.

Description

{FOOD COMPOSITION AND COSMETIC COMPOSITION FOR SKIN ANTI-AGING AND MOISTURIZING}

It relates to a food composition and a cosmetic composition for skin anti-aging and moisturizing.

The skin protects the internal organs from external environmental stimuli and plays an important role in maintaining homeostasis such as body temperature regulation. The skin ages due to various internal and external factors, and is largely divided into intrinsic aging due to genetic causes and extrinsic aging due to environmental causes. Among these, photoaging refers to aging caused by ultraviolet (UV) rays. Recently, the destruction of the ozone layer due to environmental pollution has increased the amount of UV rays, and research on photoaging has been focused on (Dermatology and Therapy, 23(1): 31-47, 2010). In photoaged skin, external features such as roughness, loss of elasticity, and wrinkles are observed, and the main research area on photoaging is changes in skin wrinkles. Many studies have reported on the basic physiological metabolic changes, such as the synthesis, decomposition, and moisture content of collagen, a major component of the skin, in relation to the formation of wrinkles on the skin due to photoaging (Annals of the New York Academy of Sciences, 973: 31-43, 2002).

Collagen, which exists in the dermis layer of the skin, accounts for approximately 70% to 80% of the total dry weight of the skin and is known to control the elasticity of the skin together with elastin, an elastic fiber. In particular, the production of reactive oxygen species increases due to ultraviolet rays and the enzymatic and non-enzymatic antioxidant defense systems of the skin are destroyed, which increases the decomposition of collagen and decreases its biosynthesis, resulting in a significant decrease in collagen in the dermis layer (Journal of Investigative Dermatology, 126(12): 2565-2575, 2006). Matrix metalloproteinases (MMPs) have a significant effect on the decrease in collagen, which are involved in the decomposition of the extracellular matrix and basement membrane. The above enzymes are activated by ultraviolet rays, and research results have been reported that inhibiting them reduces skin thickness increase and wrinkle formation induced by ultraviolet rays (Journal of Investigative Dermatology, 120(1): 128-134, 2003). Therefore, controlling Matrix Metalloproteinases (MMPs) is an effective method for preventing and treating photoaging.

The stratum corneum of the skin is the outermost layer of the skin and is in direct contact with the external environment, and thus plays an important role in protecting our bodies from external physical and chemical stresses. This barrier function is maintained by epidermal homeostasis. Epidermal homeostasis is the process of forming a skin barrier called the stratum corneum through terminal differentiation through the growth and division of keratinocytes in the basal layer and differentiation according to cell migration, thereby maintaining a continuous skin barrier function (Korean Journal of Food Science and Technology, 43: 458-463, 2011).

As keratinocytes differentiate, they produce two factors that affect hydration. First, during keratinocyte differentiation, their cell membrane is replaced by a structure called the cornified envelope. The cornified envelope is a membrane structure in which several structural proteins, including loricrin (LOR), involucrin (INV), and filaggrin (FLG), are cross-linked by enzymes called transglutaminases (TGM), which provide skin protection against the external environment and suppress moisture evaporation within the cornified envelope. Since the cornified envelope structural proteins and TGM begin to be expressed as keratinocytes differentiate, they are used as differentiation markers (Nature Reviews Molecular Cell Biology, 6(4): 328-340, 2005). Therefore, the cornified envelope and differentiation factors are used as indicators of hydration. In addition, during the differentiation process of keratinocytes, keratinocytes produce natural moisturizing factor (NMF) and maintain the function of skin barrier. The protein that is an important source of the production of NMF is FLG, and FLG is decomposed into hydrophilic amino acids by caspase 14 to form NMF. NMF maintains the moisturizing power of the skin by providing water holding capacity and moisture absorption in the air (Journal of Cell Science. 122: 1285-1294, 2009). Therefore, maintaining an appropriate level of NMF in the skin is a very important factor for skin health through the skin barrier function.

One embodiment provides a skin anti-aging and moisturizing food composition having excellent skin anti-aging activity and improving skin moisturizing function.

Another embodiment provides a skin anti-aging and moisturizing cosmetic composition having excellent skin anti-aging activity and improving skin moisturizing function.

One embodiment provides a skin anti-aging and moisturizing food composition comprising a glycine-phenylalanine derivative as an active ingredient.

The above glycine-phenylalanine derivative can be represented by the following chemical formula 1.

[Chemical Formula 1]

(In the above chemical formula 1,

R ¹ to R ⁷ are each independently hydrogen, a substituted or unsubstituted C1 to C10 alkyl group, a substituted or unsubstituted C3 to C20 cycloalkyl group, or a substituted or unsubstituted C6 to C30 aryl group,

n is 1 to 5.)

The above glycine-phenylalanine derivative may be a dipeptide compound obtained by synthesis of glycine and phenylalanine.

The above glycine-phenylalanine derivative may be included at a concentration of 1 μM to 500 μM.

The above glycine-phenylalanine derivative may be included in an amount of 0.01 wt% to 100 wt% based on the total amount of the skin anti-aging and moisturizing food composition.

The above skin anti-aging and moisturizing food composition may be a health functional food composition.

Another embodiment provides a cosmetic composition for skin anti-aging and moisturizing comprising the glycine-phenylalanine derivative as an active ingredient.

The above glycine-phenylalanine derivative may be included in an amount of 0.001 wt% to 10 wt% based on the total amount of the skin anti-aging and moisturizing cosmetic composition.

According to one embodiment, a novel use of a glycine-phenylalanine derivative is provided. That is, since the glycine-phenylalanine derivative has excellent skin anti-aging activity and improves skin moisturizing function, it can be usefully used both in a food composition for skin anti-aging and moisturizing, and also in a cosmetic composition for skin anti-aging and moisturizing.

Figure 1 is a graph showing the increase in collagen expression of a glycine-phenylalanine derivative according to Example 1 compared to Comparative Examples 1 to 3.
Figure 2 is a graph showing the inhibition of collagenase expression by a glycine-phenylalanine derivative according to Example 1 compared to Comparative Examples 1 to 3.
Figure 3 is a graph showing the promotion of differentiation of keratinocytes by glycine-phenylalanine derivatives according to Examples 1 and 2.
Figure 4 is a graph showing the increase in the expression of factors involved in promoting differentiation of keratinocytes by a glycine-phenylalanine derivative according to Example 1, compared to Comparative Examples 1 to 3.

Below, implementation examples are described in detail so that those with ordinary skill in the art can easily implement them. However, the implementation examples can be implemented in various different forms and are not limited to the implementation examples described herein.

According to one embodiment, a food composition for skin anti-aging and moisturizing containing a glycine-phenylalanine derivative as an effective ingredient is provided.

The above glycine-phenylalanine derivative may be a dipeptide compound obtained by synthesis of two amino acids, namely, glycine and phenylalanine, and may be specifically represented by the following chemical formula 1.

[Chemical Formula 1]

(In the above chemical formula 1,

R ¹ to R ⁷ are each independently hydrogen, a substituted or unsubstituted C1 to C10 alkyl group, a substituted or unsubstituted C3 to C20 cycloalkyl group, or a substituted or unsubstituted C6 to C30 aryl group,

n is 1 to 5.)

According to one embodiment, the glycine-phenylalanine derivative inhibits collagen decomposition enzymes and promotes collagen production, so it has excellent skin anti-aging activity and can be used as an effective ingredient in food compositions and cosmetic compositions. In addition, the glycine-phenylalanine derivative promotes differentiation of keratinocytes and thus has excellent skin moisturizing function and can be used as an effective ingredient in food compositions and cosmetic compositions.

The above glycine-phenylalanine derivative can be included at a concentration of 1 μM to 500 μM, for example, can be included at a concentration of 100 μM to 400 μM. When the glycine-phenylalanine derivative is included in the above concentration range, it can enhance skin anti-aging activity by inhibiting collagen decomposition enzyme expression and increasing collagen expression, and can also enhance the ability to inhibit moisture loss by promoting differentiation of keratinocytes.

The above glycine-phenylalanine derivative may be included in an amount of 0.01 wt% to 100 wt% based on the total amount of the skin anti-aging and moisturizing food composition, for example, 0.02 wt% to 99 wt%. When the glycine-phenylalanine derivative is included within the above content range, it can increase skin anti-aging activity and improve skin moisturizing function, and has excellent safety and is easy to manufacture in a formulation.

The above-mentioned skin anti-aging and moisturizing food composition may include all forms such as functional food, nutritional supplement, health functional food, food additive, feed, etc., and is intended for consumption by humans or animals including livestock. The above-mentioned type of food composition may be manufactured in various forms according to conventional methods known in the art.

For example, as general foods, the glycine-phenylalanine can be added to, but is not particularly limited to, beverages including alcoholic beverages, fruits and processed foods thereof (e.g., canned fruits, bottled fruits, jams, marmalades, etc.), fish, meats and processed foods thereof (e.g., ham, sausages, corned beef, etc.), breads and noodles (e.g., udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, taffy, dairy products (e.g., butter, cheese, etc.), edible plant fats, margarine, vegetable proteins, retort foods, frozen foods, various seasonings (e.g., soybean paste, soy sauce, sauces, etc.), and the like, and manufactured. In addition, as a nutritional supplement, the glycine-phenylalanine can be added to, but is not particularly limited to, capsules, tablets, pills, etc. and manufactured. In addition, as a health functional food, although not particularly limited, the glycine-phenylalanine itself can be manufactured in the form of tea, juice, and drink, and can be consumed by liquefying, granulating, encapsulating, and powdering so that it can be consumed (health drink). In addition, in order to use the glycine-phenylalanine in the form of a food additive, it can be manufactured in the form of a powder or concentrate and used. In addition, the glycine-phenylalanine can be manufactured in the form of a composition by mixing it with a known active ingredient known to have skin anti-aging and moisturizing improvement effects.

When the above-mentioned skin anti-aging and moisturizing food composition is used as a health beverage composition, it may contain various flavoring agents or natural carbohydrates as additional ingredients like a regular beverage. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; sugar alcohols such as xylitol, sorbitol and erythritol. The sweetener may be a natural sweetener such as thaumatin and stevia extract; a synthetic sweetener such as saccharin and aspartame, etc.

The above glycine-phenylalanine may also be contained as an effective ingredient of a food composition for the prevention or treatment of a disease mediated by skin anti-aging and moisturizing improvement, and the amount thereof is not particularly limited to an amount effective to achieve the effect of preventing or treating a disease mediated by skin anti-aging and moisturizing improvement, but may be 0.01 wt% to 100 wt% based on the total amount of the entire composition.

A food composition for skin anti-aging and moisturizing according to one embodiment can be prepared by mixing glycine-phenylalanine with other active ingredients known to improve skin anti-aging and moisturizing properties.

When the above skin anti-aging and moisturizing food composition is used as a health functional food composition, it may further contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents, etc. In addition, it may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, or vegetable drinks. These added ingredients may be used independently or in combination.

According to another embodiment, a cosmetic composition for skin anti-aging and moisturizing comprising a glycine-phenylalanine derivative as an active ingredient is provided.

Since the glycine-phenylalanine derivative used as an effective ingredient in the above cosmetic composition is the same as that used as an effective ingredient in the aforementioned food composition, the same description is omitted.

The above glycine-phenylalanine derivative may be included in an amount of 0.001 wt% to 10 wt% based on the total amount of the skin anti-aging and moisturizing cosmetic composition, for example, 0.01 wt% to 5 wt%. When the glycine-phenylalanine derivative is included within the above content range, it can increase skin anti-aging activity and improve skin moisturizing function, and has excellent safety and is easy to manufacture in a formulation.

A cosmetic composition for skin anti-aging and moisturizing according to one embodiment may further comprise an additional effective ingredient.

The above additional effective ingredients may be known skin anti-aging and moisturizing improvement ingredients, and examples thereof include hyaluronic acid, vitamin C, niacinamide, retinoic acid, or a combination thereof.

When including the above additional effective ingredients, skin safety, ease of formulation, and stability of the effective ingredients according to combined use with the above glycine-phenylalanine derivative can be considered.

The additional effective ingredient may be included in an amount of 0.0001 wt% to 10 wt% based on the total amount of the skin anti-aging and moisturizing cosmetic composition, for example, 0.0001 wt% to 1 wt%, 0.0001 wt% to 0.1 wt%, 0.0001 wt% to 0.001 wt%, 0.001 wt% to 10 wt%, 0.001 wt% to 1 wt%, 0.001 wt% to 0.1 wt%, 0.01 wt% to 10 wt%, 0.01 wt% to 1 wt%. The content range may be adjusted depending on requirements such as skin safety and ease of formulation.

According to one embodiment, a cosmetic composition for skin anti-aging and moisturizing comprises the glycine-phenylalanine derivative as an effective ingredient and further comprises a dermatologically acceptable excipient, thereby being manufactured in the form of a basic cosmetic composition such as a cleanser, such as a toner, a cream, an essence, a cleansing foam and a cleansing water; a pack, a body oil; a color cosmetic composition such as a foundation, a lipstick, a mascara and a makeup base; a hair product composition such as a shampoo, a rinse, a hair conditioner and a hair gel; and a soap.

The above excipients are not particularly limited, but may include, for example, skin softeners, skin penetration enhancers, colorants, fragrances, emulsifiers, thickeners, solvents, fragrances, pigments, sterilizers, antioxidants, preservatives, moisturizers, etc. In addition, thickeners, inorganic salts, synthetic polymers, etc. may be included for the purpose of improving physical properties.

For example, when manufacturing a cleanser and soap with the skin anti-aging and moisturizing cosmetic composition according to one embodiment, the cleanser and soap can be easily manufactured by adding the glycine-phenylalanine derivative to a typical cleanser and soap base. As another example, when manufacturing a cream, the cream can be manufactured by adding the glycine-phenylalanine derivative or a salt thereof to a typical oil-in-water (O/W) cream base. Here, synthetic or natural materials such as fragrances, chelating agents, pigments, antioxidants, preservatives, and proteins, minerals, and vitamins for the purpose of improving physical properties can be additionally added.

Hereinafter, specific examples of the present invention will be presented. However, the examples described below are only intended to specifically illustrate or explain the present invention, and the present invention should not be limited thereto. In addition, since the contents not described herein can be sufficiently technically inferred by those skilled in the art, their descriptions will be omitted.

(glycine-phenylalanine derivative)

The glycine-phenylalanine derivative used in Examples 1 and 2 below is a compound represented by the following chemical formula 2.

[Chemical formula 2]

Molecular weight: 222.24 g/mol

Where to buy: Sigma-Aldrich (Saint Louis, MO, USA)

Experimental Example 1: Increased Expression of Collagen COL1A1 by Glycine-Phenylalanine Derivatives

Hs68 human dermal fibroblasts (ATCC, Manassas, VA, USA) were seeded at 2 × 10 5 cells/well in 6-well plates with Dulbecco's modified Eagle's media (DMEM; Hyclone) containing 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA). After 90% confluency, the medium was removed, washed twice with phosphate buffered saline (PBS), and 1 mL of PBS was added per well. Then, the plates were irradiated with ¹⁵ mJ/cm ² of UV light. After treating the plate with 15 mJ/cm ² of UV rays, the PBS was removed and DMEM (Hyclone) containing 300 μM glycine alone (Comparative Example 1), 300 μM phenylalanine alone (Comparative Example 2), 300 μM of a mixture of glycine and phenylalanine (Comparative Example 3), and 300 μM glycine-phenylalanine (Sigma-Aldrich, G2752) (Example 1) was treated for 24 hours. At this time, the case where only DMEM was treated without anything was used as the control. The isolated total RNA was quantified using NanoDrop 1000; Thermo Fisher Scientific Inc. Waltham, MA, USA). The quantified 16 μL of RNA was used to synthesize cDNA using 4 μL of reverse transcriptase premix (ELPIS-Biotech, Daejeon, Korea) and a PCR machine (T100 Thermal Cycler; Bio-rad, Hercules, CA, USA) at 42°C for 60 min and 94°C for 5 min.

1 μL of cDNA, the following specific primers (Bioneer, Daejeon, Korea), and qPCR 2X Premix (Dyne bio, Seongnam, Korea) were used, and the reaction was performed at 95 °C for 10 min, followed by 50 cycles of 95 °C for 10 s, 60 °C for 30 s, and 72 °C for 20 s, and then 72 °C for 5 min. The loading amount of mRNA was constant with GAPDH.

COL1A1

Forward primer: 5’-CACGACAAAGCAGAAACATC- 3’ (SEQ ID NO: 1)

Reverse primer: 5’-ACACATCAAGACAAGAACGAG- 3’ (SEQ ID NO: 2)

GAPDH

Forward primer: 5’-TCACCACCATGGAGAAGGC- 3’ (SEQ ID NO: 1)

Reverse primer: 5’-GCTAAGCAGTTGGTGGTGCA - 3’ (SEQ ID NO: 2)

qPCR results were obtained using CFX94 Maestro Software, and the results were quantified with GAPDH and shown in Figure 1.

Figure 1 is a graph showing the increase in collagen expression of a glycine-phenylalanine derivative according to Example 1 compared to Comparative Examples 1 to 3.

Through Fig. 1, it can be confirmed that the mRNA expression of COL1A1 decreased by ultraviolet B (UVB) was increased the most in Hs68 human skin fibroblasts by treating with the glycine-phenylalanine derivative according to Example 1 compared to Comparative Examples 1 to 3. However, it can be seen that there was no effect, especially when glycine and phenylalanine were mixed and treated according to Comparative Example 3. From this, it can be seen that the glycine-phenylalanine derivative according to one embodiment has excellent skin anti-photoaging ability by increasing collagen expression in skin fibroblasts.

Experimental Example 2: Inhibition of collagen-degrading enzyme MMP-1 expression by glycine-phenylalanine derivatives

Hs68 human dermal fibroblasts (ATCC, Manassas, VA, USA) were seeded at 2 × 10 5 cells/well in 6-well plates with Dulbecco's modified Eagle's media (DMEM; Hyclone) containing 10% fetal bovine serum (FBS; Hyclone, Logan, UT, USA). After 90% confluency, the medium was removed, washed twice with phosphate buffered saline (PBS), 1 mL of PBS was added per well, and then the plates were irradiated with ¹⁵ mJ/cm ² of UV light. After 15 mJ/cm ² of UV light was treated to the plates, the PBS was removed, and DMEM (Hyclone) containing 300 μM of a glycine-phenylalanine derivative (Example 1) was treated for 24 h. At this time, the case where only DMEM was treated without anything was used as the control. The isolated total RNA was quantified using NanoDrop 1000; Thermo Fisher Scientific Inc. Waltham, MA, USA). The quantified 16 μL RNA was used to synthesize cDNA using 4 μL of reverse transcriptase premix (Reverse Transcriptase Premix; ELPIS-Biotech, Daejeon, Korea) in a PCR machine (T100 Thermal Cycler; Bio-rad, Hercules, CA, USA) at 42°C for 60 min and 94°C for 5 min.

1 μL of cDNA, the following specific primers (Bioneer, Daejeon, Korea), and qPCR 2X Premix (Dyne bio, Seongnam, Korea) were used at 95 °C for 10 min, followed by 50 cycles of 95 °C for 10 s, 60 °C for 30 s, and 72 °C for 20 s, and then 72 °C for 5 min. The loading amount of mRNA was constant with GAPDH.

MMP-1

Forward primer: 5’-AAGTCAAGTTTGTGGCTTATGG- 3’ (SEQ ID NO: 1)

Reverse primer: 5’-GACTCATGTCTCCTGTCTCTTTCT- 3’ (SEQ ID NO: 2)

GAPDH

Forward primer: 5’-TCACCACCATGGAGAAGGC- 3’ (SEQ ID NO: 1)

Reverse primer: 5’-GCTAAGCAGTTGGTGGTGCA- 3’ (SEQ ID NO: 2)

qPCR results were obtained using CFX94 Maestro Software, and the results were quantified with GAPDH and shown in Figure 2.

Figure 2 is a graph showing the inhibition of collagenase expression by a glycine-phenylalanine derivative according to Example 1.

Through Fig. 2, it can be confirmed that the mRNA expression of MMP-1 increased by ultraviolet B (UVB) was reduced in Hs68 human dermal fibroblasts by treating with the glycine-phenylalanine derivative according to Example 1. From this, it can be seen that the glycine-phenylalanine derivative according to one embodiment has an excellent ability to improve skin wrinkles by inhibiting the expression of collagen-decomposing enzyme in dermal fibroblasts.

Experimental Example 3: Keratinogenesis-promoting activity by glycine-phenylalanine derivatives

HaCaT human keratinocytes (ATCC) were placed in 6-well plates with DMEM (Hyclone) containing 10% FBS (Hyclone). After growing to 100% confluency, the medium was removed, and glycine-phenylalanine derivatives were dissolved in DMEM (Hyclone) to 300 μM (Example 1) and glycine-phenylalanine derivatives (Example 2), and then treated at various concentrations. At this time, the group treated with DMEM containing nothing instead of glycine-phenylalanine derivatives was used as the control group. After 72 hours, NP-40 buffer (ELPIS-Biotech, Daejeon) containing 2% sodium dodecyl sulfate (SDS) was treated. After harvesting the cells, they were centrifuged at 14,000 rpm for 10 minutes, and the supernatant was removed. To dissolve the precipitate, distilled water containing 2% SDS and 20 mM dithiothreitol (DTT) was added and boiled at 100°C for approximately 60 minutes. After 60 minutes, the supernatant was collected and the absorbance was measured at a wavelength of 340 nm and compared, and the results are shown in Fig. 3.

Figure 3 is a graph showing the promotion of differentiation of keratinocytes by glycine-phenylalanine according to Examples 1 and 2.

Through Fig. 3, it can be confirmed that differentiation of HaCaT human keratinocytes was promoted by treatment with glycine-phenylalanine according to Examples 1 and 2. From this, it can be seen that the glycine-phenylalanine derivative according to one embodiment has an excellent ability to promote differentiation of keratinocytes and suppress water loss in keratinocytes.

Experimental Example 4: Increased activity of keratinogenesis-promoting factor expression by glycine-phenylalanine derivatives

HaCaT human keratinocytes (ATCC) were placed in a 6-well plate with DMEM (Hyclone) containing 10% FBS (Hyclone). After growing to 100% confluency, the medium was removed, and DMEM (Hyclone) containing 300 μM glycine alone (Comparative Example 1), 300 μM phenylalanine alone (Comparative Example 2), 300 μM of a mixture of glycine and phenylalanine (Comparative Example 3), and 300 μM glycine-phenylalanine (Sigma-Aldrich, G2752) (Example 1) was treated for 24 hours. At this time, the case treated only with DMEM containing nothing was used as the control. The isolated total RNA was quantified using NanoDrop 1000; Thermo Fisher Scientific Inc. Waltham, MA, USA). The quantified 16 μL of RNA was used to synthesize cDNA using 4 μL of reverse transcriptase premix (ELPIS-Biotech, Daejeon, Korea) and a PCR machine (T100 Thermal Cycler; Bio-rad, Hercules, CA, USA) at 42°C for 60 min and 94°C for 5 min.

1 μL of cDNA, the following specific primers (Bioneer, Daejeon, Korea), and qPCR 2X Premix (Dyne bio, Seongnam, Korea) were used, and the reaction was performed at 95 °C for 10 min, followed by 50 cycles of 95 °C for 10 s, 60 °C for 30 s, and 72 °C for 20 s, and then 72 °C for 5 min. The loading amount of mRNA was constant with GAPDH.

Filaggrin

Forward primer: 5’- TGAGGGCACTGAAAGGCAAA- 3’ (SEQ ID NO: 1)

Reverse primer: 5’- TGGCCACATAAACCTGGGTC- 3’ (SEQ ID NO: 2)

Loricrin

Forward primer: 5’- GGGTACCACGGAGGCGAAGGA- 3’ (SEQ ID NO: 1)

Reverse primer: 5’- ACTGAGGCACTGGGGTTGGGA- 3’ (SEQ ID NO: 2)

Transglutaminase

Forward primer: 5’- GACAATGGTGTGCTTGCTGG- 3’ (SEQ ID NO: 1)

Reverse primer: 5’- TGCACATCACCCTCTCGAAC- 3’ (SEQ ID NO: 2)

Caspase 14

Forward primer: 5’- CGGGACTCACAACCAAAGGA- 3’ (SEQ ID NO: 1)

Reverse primer: 5’- GGGTCCCTTTGTTCTCCTCG- 3’ (SEQ ID NO: 2)

GAPDH

Forward primer: 5’-TCACCACCATGGAGAAGGC- 3’ (SEQ ID NO: 1)

Reverse primer: 5’-GCTAAGCAGTTGGTGGTGCA - 3’ (SEQ ID NO: 2)

qPCR results were obtained using CFX94 Maestro Software, and the results were quantified with GAPDH and shown in Figure 4.

Figure 4 is a graph showing the increase in the expression of factors involved in promoting differentiation of keratinocytes by a glycine-phenylalanine derivative according to Example 1, compared to Comparative Examples 1 to 3.

Through FIG. 4, it can be confirmed that the mRNA expression of Filaggrin, Loricrin, Transglutaminase, and Caspase 14 increased the most in HaCaT human keratinocytes by treating with glycine-phenylalanine according to Example 1 compared to Comparative Examples 1 to 3. In particular, when glycine and phenylalanine were mixed and treated according to Comparative Example 3, an effect was shown compared to the control group, but it can be seen that the effect of the glycine-phenylalanine derivative was more excellent compared to the glycine-phenylalanine according to Example 1. From this, it can be seen that the glycine-phenylalanine derivative according to one embodiment has an excellent skin barrier strengthening ability by promoting keratinogenesis by increasing the expression of Filaggrin, Loricrin, Transglutaminase, and Caspase 14 in human keratinocytes.

(Manufacture of food composition for skin anti-aging and moisturizing)

Manufacturing Example 1-1: Health Functional Food

According to one embodiment, 1000 mg of glycine-phenylalanine (Sigma-Aldrich, G2752), 70 ug of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B1, 0.15 mg of vitamin B2, 0.5 mg of vitamin B6, 0.2 ug of vitamin B12, 10 mg of vitamin C, 10 ug of biotin, 1.7 mg of nicotinamide, 50 ug of folic acid, 0.5 mg of calcium pantothenate, 1.75 mg of ferrous sulfate, 0.82 mg of zinc oxide, 25.3 mg of magnesium carbonate, 15 mg of monobasic potassium phosphate, 55 mg of dibasic calcium phosphate, 90 mg of potassium citrate, 100 mg of calcium carbonate, and 24.8 mg of magnesium chloride were mixed to prepare. At this time, the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method, granules are manufactured, and a health functional food composition is manufactured according to a conventional method.

Manufacturing Example 1-2: Health Drink

According to one embodiment, 1000 mg of glycine-phenylalanine (Sigma-Aldrich, G2752), 1000 mg of citric acid, 100 g of oligosaccharide, 2 g of plum concentrate, and 1 g of taurine were mixed with purified water to make a total of 900 mL according to a conventional health beverage manufacturing method, and then the above ingredients were mixed and heated at 85°C for about 1 hour, and the resulting solution was filtered, placed in a sterilized 2 L container, sealed and sterilized, and then stored in the refrigerator to manufacture a health beverage composition.

Manufacturing Example 1-3: Chewing gum

Chewing gum was manufactured by a conventional method by mixing 20 wt% of gum base, 76.9 wt% of sugar, 3 wt% of water, and 0.1 wt% of glycine-phenylalanine (Sigma-Aldrich, G2752) according to one embodiment.

Manufacturing Example 1-4: Candy

Candy was manufactured by a conventional method by mixing 60 wt% of sugar, 39.9 wt% of corn syrup, and 0.1 wt% of glycine-phenylalanine (Sigma-Aldrich, G2752) according to one embodiment.

Manufacturing Example 1-5: Biscuit

25.59 wt% of 1st class strength, 22.22 wt% of 1st gravity strength, 4.80 wt% of refined sugar, 0.73 wt% of salt, 0.78 wt% of glucose, 11.78 wt% of palm shortening, 1.54 wt% of ammonium, 0.17 wt% of baking soda, 0.16 wt% of sodium bisulfite, 1.45 wt% of rice flour, 0.0001 wt% of vitamin B, 0.04 wt% of milk flavor, 20.6998 wt% of water, 1.16 wt% of whole milk powder, 0.29 wt% of milk substitute, 0.03 wt% of monobasic calcium phosphate, 0.29 wt% of spraying salt, 7.27 wt% of spraying oil, and 1 wt% of glycine-phenylalanine (Sigma-Aldrich, G2752) according to one embodiment. Biscuits were made by mixing the ingredients in the usual way.

(Manufacture of cosmetic composition for skin anti-aging and moisturizing)

Manufacturing Example 2-1: Nutritious Toner (Milk Lotion)

According to an embodiment of the present invention, a nutritional toner was prepared by a conventional method according to the nutritional toner formulation ratio of Table 1 below using glycine-phenylalanine (Sigma-Aldrich, G2752). At this time, the total amount of the ingredients is 100.

Mixing ingredients Content (weight%) Glycine-phenylalanine derivatives 2.0 Squalane 5.0 beeswax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.5 Liquid paraffin 0.5 Caprylic/Capric Triglyceride 5.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Triethanolamine 0.2 Purified water Remaining amount

Manufacturing Example 2-2: Flexible Toner (Skin Lotion)

According to an embodiment of the present invention, a flexible toner was manufactured by a conventional method according to the formulation ratio of the flexible toner in Table 2 below using glycine-phenylalanine (Sigma-Aldrich, G2752). At this time, the total amount of the ingredients is 100.

Mixing ingredients Content (weight%) Glycine-phenylalanine derivatives 2.0 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 PEG 12 nonylphenyl ether 0.2 Polysorbate 80 0.4 Ethanol 10.0 Triethanolamine 0.1 Purified water Remaining amount

Manufacturing Example 2-3: Nourishing Cream

According to an embodiment of the present invention, a nutritional cream was prepared by a conventional method according to the nutritional cream formulation ratio in Table 3 below using glycine-phenylalanine (Sigma-Aldrich, G2752). At this time, the total amount of the ingredients is 100.

Mixing ingredients Content (weight%) Glycine-phenylalanine derivatives 2.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 PEG60 hydrogenated castor oil 2.0 Liquid paraffin 10 Squalane 5.0 Caprylic/Capric Triglyceride 5.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Purified water Remaining amount

Manufacturing Example 2-4: Massage Cream

According to an embodiment of the present invention, a massage cream was prepared by a conventional method according to the massage cream formulation ratio in Table 4 below using glycine-phenylalanine (Sigma-Aldrich, G2752). At this time, the total amount of the ingredients is 100.

Mixing ingredients Content (weight%) Glycine-phenylalanine derivatives 1.0 beeswax 10.0 Polysorbate 60 1.5 PEG 60 hydrogenated castor oil 2.0 Sorbitan sesquioleate 0.8 Liquid paraffin 40.0 Squalane 5.0 Caprylic/Capric Triglyceride 4.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Purified water Remaining amount

Manufacturing Example 2-5: Pack

According to an embodiment of the present invention, a pack was manufactured by a conventional method according to the pack formulation ratio of Table 5 below using glycine-phenylalanine (Sigma-Aldrich, G2752). At this time, the total amount of the components is 100.

Mixing ingredients Content (weight%) Glycine-phenylalanine derivatives 1.0 polyvinyl alcohol 13.0 Sodium carboxymethyl cellulose 0.2 glycerin 5.0 Allantoin 0.1 Ethanol 6.0 PEG 12 nonylphenyl ether 0.3 Polysorbate 60 0.3 Purified water Remaining amount

Manufacturing Example 2-6: Gel

According to an embodiment of the present invention, a gel was prepared by a conventional method according to the gel formulation ratio of Table 6 below using glycine-phenylalanine (Sigma-Aldrich, G2752). At this time, the total amount of the components was 100.

Mixing ingredients Content (weight%) Glycine-phenylalanine derivatives 0.5 Sodium ethylenediamine acetate 0.05 glycerin 5.0 Carboxyvinyl polymer 0.3 Ethanol 5.0 PEG 60 hydrogenated castor oil 0.5 Triethanolamine 0.3 Purified water Remaining amount

Although the preferred embodiments of the present invention have been described above, the present invention is not limited thereto, and various modifications may be made within the scope of the claims, the detailed description of the invention, and the attached drawings, which also fall within the scope of the present invention.

Claims (11)

A food composition for skin anti-aging and moisturizing comprising a glycine-phenylalanine derivative represented by the following chemical formula 2 as an effective ingredient:
[Chemical formula 2]
. delete In paragraph 1,
The above glycine-phenylalanine derivative is a skin anti-aging and moisturizing food composition which is a dipeptide compound obtained by the synthesis of glycine and phenylalanine. In paragraph 1,
A food composition for skin anti-aging and moisturizing, wherein the glycine-phenylalanine derivative is contained in a concentration of 1 μM to 500 μM. In paragraph 1,
A skin anti-aging and moisturizing food composition, wherein the glycine-phenylalanine derivative is contained in an amount of 0.01 wt% to 100 wt% based on the total amount of the skin anti-aging and moisturizing food composition. In paragraph 1,
The above skin anti-aging and moisturizing food composition is a skin anti-aging and moisturizing food composition that is a health functional food composition. A cosmetic composition for skin anti-aging and moisturizing comprising a glycine-phenylalanine derivative represented by the following chemical formula 2 as an effective ingredient:
[Chemical formula 2]
. delete In Article 7,
The above glycine-phenylalanine derivative is a skin anti-aging and moisturizing cosmetic composition which is a dipeptide compound obtained by synthesis of glycine and phenylalanine. In Article 7,
A cosmetic composition for skin anti-aging and moisturizing, wherein the glycine-phenylalanine derivative is contained in a concentration of 1 μM to 500 μM. In Article 7,
A skin anti-aging and moisturizing cosmetic composition, wherein the glycine-phenylalanine derivative is contained in an amount of 0.001 wt% to 10 wt% based on the total amount of the skin anti-aging and moisturizing cosmetic composition.