KR102193209B1 - Cosmetic composition for skin whitening comprising extract of Phragmites Communis - Google Patents

Abstract

The present invention relates to a skin-whitening cosmetic composition including a hot water extract of Phragmites communis as an active ingredient, a manufacturing method thereof, and a quasi-drug for skin whitening including a high temperature and pressurized hot water extract of Phragmites communis as an active ingredient. The manufacturing method comprises the following steps: (a) extracting reeds (Phragmites communis) with hot water; (b) cooling the extract; and (c) acquiring the cooled extract as an extract in the form of powder. According to the present invention, the composition includes a reed root extract extracted by applying high temperature and pressure as an active ingredient, and the extract has an antioxidant effect, inhibits tyrosinase and alpha-glucosidase activities, and inhibits the activity of melanocytes, thereby being used for skin-whitening to prevent and mitigate pigmentation of the skin.

Description

[Cosmetic composition for skin whitening comprising extract of Phragmites Communis}

The present invention is a cosmetic composition for skin whitening comprising a hot water extract of reeds (Phragmites Communis) as an active ingredient, (a) extracting hot water of reeds (Phragmites Communis); (b) cooling the extract; And (c) a method for preparing a cosmetic composition for skin whitening comprising the step of obtaining the cooled extract as an extract in the form of a powder, and a quasi-drug composition for skin whitening comprising a hot and pressurized hot water extract of Phragmites Communis as an active ingredient. About.

Human skin color is genetically determined according to the concentration and distribution of melanin in the skin, but is also affected by environmental or physiological conditions such as solar ultraviolet rays, fatigue, and stress.

The chemical action on the production of melanin, which determines the color of the skin, has been reported in a number of papers so far.Melanocytes, which are melanocytes, protect cells from ultraviolet rays in the intracellular organelle called melanosomes. After synthesizing melanin having a protective function, it is known that the melanin migrates from melanocytes to keratinocytes, and it is reported that the melanin transferred to the keratinocytes determines the skin color.

It is known that the precursor required in the melanin biosynthesis process is an amino acid called tyrosine, and this tyrosine is converted to dihydroxyphenylalanine (DOPA), and then finally converted to melanin through a substance called dopaquinone. .

In this regard, melanin-forming enzymes include tyrosinase (TYR), tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (tyrosinase-related protein). 2, TRP-2/DOPA, chrome tautomerase) and the like are known, and the melanin-forming enzyme is known to be regulated by a transcription factor called a microphthalmia-associated transcription factor (MITF). Tyrosinase, which plays an important role in the melanin synthesis pathway, converts L-tyrosine to L-DOPA (L-3,4-dihydroxyphenylalanine), and rapidly converts the generated L-DOPA to L-dopaquinone, Through subsequent reactions, it eventually forms melanin. Melanin synthesized by tyrosinase darkens the skin color, and research on a skin whitening method to brighten the skin color by inhibiting the melanin synthesis due to health or beauty requirements is actively underway.

As a commonly known whitening ingredient, arbutin, kojic acid (kojic acid), azelaic acid, aloesin, 4-butylresorcinol, resveratrol, ceramide, sphingosine-1-phosphate, sphingosyl which inhibit tyrosinase enzyme activity There are substances such as phosphorylcholine and the like, and hydroquinone (Korean Patent Application Publication No. 2016-0014271). These whitening components inhibit the synthesis of melanin pigment, thereby brightening the skin tone to realize skin whitening, or to improve skin hyperpigmentation such as spots and freckles caused by ultraviolet rays, hormones, or genetics. However, when some whitening ingredients are applied to the skin, there is a problem in that the amount of use is limited due to safety problems such as irritation and redness, or the effect is insignificant and thus no practical effect can be expected. In particular, in the case of hydroquinone, its use was prohibited due to unwanted side effects such as irreversible bleaching (European Committee 24th Dir 2000/6/EC), and it was replaced with arbutin, a structural analog of hydroquinone. As described above, in the field of whitening, it is particularly important to not only have a whitening effect by having an effect of inhibiting tyrosinase activity, but also to secure stability, and development of a material that satisfies all of the above requirements is still required.

In recent years, along with lifestyle changes and interest in well-being, the demand for well-being is gradually increasing. Accordingly, the interest of consumers is increasing in the field of skin beauty as well, and related skin whitening products are being developed in various ways, but the reliability of actual consumers in the efficacy is not very high. In addition, as the wind of naturalism has recently increased, preference for products made of natural materials is increasing, and the cosmetic market, which is based on technology using natural materials, is rapidly increasing. Manufacturing cosmetics using natural ingredients has the advantage of improving the absorption rate and safety of active ingredients of natural ingredients.

Accordingly, the inventors of the present invention made diligent efforts to develop a material that can substantially help skin whitening effect while securing safety when applied to the skin by using natural materials as a material for cosmetics with a skin whitening function. The extract obtained by extracting the roots at high temperature and pressurization has more than 5 times higher content of coumarin acid than general hot water extract, has excellent L-DOPA oxidation inhibitory ability, has high antioxidant efficacy even compared to ethanol (70%, 100%) extract, and tyrosinase The present invention was completed by discovering for the first time that it has excellent whitening effect due to its high activity inhibitory ability and α-glucosidase activity inhibitory ability, and it can be used for skin whitening purposes.

The present invention relates to a cosmetic composition for skin whitening comprising a reed hot water extract as an active ingredient.

The present invention comprises the steps of (a) extracting hot water from reeds; (b) cooling the extract; And (c) it relates to a method for producing a cosmetic composition for skin whitening comprising the step of obtaining the cooled extract as a powder extract.

The present invention relates to a quasi-drug composition for skin whitening comprising a reed hot and pressurized hot water extract as an active ingredient.

This will be described in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention can be applied to each other description and embodiment. That is, all combinations of various elements disclosed in the present invention belong to the scope of the present invention. In addition, it cannot be seen that the scope of the present invention is limited by the specific description described below.

One aspect of the present invention for achieving the above object provides a cosmetic composition for skin whitening comprising a reed hot water extract as an active ingredient.

The present invention is safe when applied to the skin when the extract extracted from the natural material reed root through high temperature and pressurization, the content of coumarin acid is 5 times higher than that of general hot water extract, L-DOPA oxidation inhibitory ability is excellent, ethanol (70% , 100%) compared to the extract, it has high antioxidant efficacy, high tyrosinase activity inhibitory ability and α-glucosidase activity inhibitory activity, so it has excellent whitening effect.Based on the technical idea to use this in skin whitening compositions As such, its efficacy was first identified by the present inventors.

In the present invention, the term "reed (Phragmites Communis)" is a perennial plant of the monocotyledonous herbaceous plant, and refers to a plant that forms a colony and grows in the sandy land around a wetland or a mandala or a lake. Specifically, in the present invention, the reed may mean the root of the reed, but is not particularly limited thereto. The root of the reed is also referred to as "nogeun", which means that the root of the reed is dried in oriental medicine. The pharmacological action of Nogeun is known for its diuretic action, antipyretic action, hepatic protection action, and hematopoietic reinforcement action.

In the present invention, the term "hot water" means hot water.

In this case, “high temperature” means a high temperature, and in the present invention, the high temperature means 110°C or more, more specifically, 120°C or more, but is not limited thereto.

In the present invention, the term "extract" refers to a liquid component obtained by immersing a target substance in various solvents and then extracting it at room temperature, low temperature or warming state for a certain period of time, and a resultant product such as a solid component obtained by removing the solvent from the liquid component. Means. In addition, it can be comprehensively interpreted as including all of the resulting diluted solution, a concentrated solution thereof, a dried product obtained by drying the extract, or a preparation thereof, a purified product, and the like.

More specifically, the extract may be hot water extraction under pressurized conditions, and the “pressurization” refers to applying pressure, and in the present invention, the pressurization may be 10 psi to 20 psi, but is not limited thereto.

The extract may be extracted without limitation from natural, hybrid, and variety plants, and may be obtained by extraction using water (purified water), C1 to C6 alcohol, and a mixed solvent thereof. In addition, the method for obtaining the extract is not particularly limited as long as the reed extract of the present invention can be obtained, but specifically, the reed, reed root, etc., are immersed in the solvent and extracted at room temperature, such as cold sediment extraction, heating Methods such as extraction, ultrasonic extraction by applying ultrasonic waves, and reflux extraction using a reflux cooler may be used.

In the present invention, the hot water extract may be an extract of reed root, specifically, may be an extract obtained through high temperature and pressure, and more specifically, 10 psi to 20 psi at a high temperature of 10 °C to 130 °C. Hot water extraction may be performed for minutes to 20 minutes, and the hot water extract may be prepared in a powder form through lyophilization, but is not limited thereto.

In the present invention, the term "skin whitening" refers to brightening skin tone by inhibiting the synthesis of melanin pigment and/or inhibiting tyrosinase activity. In addition, the skin whitening includes improving the deposition of skin hyperpigmentation such as spots, freckles, age spots, blemishes or melanoma caused by ultraviolet rays, hormones or genetics.

The melanin is a pigment of dark brown particles, and is produced by organelles called melanosomes in melanocytes. Melanin exists in the skin, hair, and eyes, and the skin color is determined by the amount of melanin. The higher the amount of melanin, the darker the skin color is.

Accordingly, the composition may be characterized by a skin whitening effect that suppresses melanin production.

In one embodiment of the present invention, it was confirmed that the reed hot water extract has high antioxidant efficacy, tyrosinase activity inhibitory ability, and α-glucosidase activity inhibitory activity compared to the reed ethanol (70%, 100%) extract. By treating the melanocytes, it was confirmed that the amount of pigment production of melanin was reduced.

Through this, it was confirmed that the composition containing the high temperature/pressurized reed extract of the present invention as an active ingredient can be used as a cosmetic composition for skin whitening.

In addition, in one embodiment of the present invention, it was confirmed that the hot water extract of reed root extracted by high temperature and pressure than the hot water extract of general reed root has a content of coumarin acid 5 times or more and has excellent L-DOPA oxidation inhibitory ability.

Through this, it was confirmed that the high temperature/pressurized reed extract of the present invention is superior to the general reed hot water extract.

The "cosmetic composition" of the present invention may contain a high temperature/pressurized reed extract in an amount of 0.0001 to 50% by weight (w/w) based on the total weight of the composition, and specifically 0.001 to 5% by weight (w/w) However, it is not particularly limited thereto. The cosmetic composition is a softening lotion, astringent lotion, nutritional lotion, nutrition cream, massage cream, essence, pack, skin patch, skin adhesion gel, powder, ointment, paste, gel, suspension, emulsion, spray, essence or capsule. It may be formulated as, but is not particularly limited thereto.

The cosmetic composition of the present invention may additionally include one or more cosmetically acceptable carriers blended in general skin cosmetics, and as common ingredients, for example, oil, water, surfactant, moisturizer, lower alcohol, thickener, A chelating agent, a colorant, a preservative, a fragrance, and the like may be appropriately blended, but the present invention is not limited thereto. The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation.

When the formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or Mixtures of these can be used.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or mixtures thereof may be used as a carrier component, and in particular, in the case of a spray, additional chloro Propellants such as fluorohydrocarbon, propane/butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil may be used, and in particular, cottonseed oil, peanut oil, corn seed oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan may be used. have.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, micro Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, and the like may be used.

Meanwhile, when the formulation of the present invention is a capsule, it may be formulated in the form of alginate capsules, agar capsules, gelatin capsules, wax capsules, or double capsules. It is not limited thereto.

In addition, the cosmetic composition of the present invention is a supplement that is commonly used in addition to the high temperature and pressurized hot water extract of reeds, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic activators, preservatives, antioxidants, solvents, fragrances, fillers, blockers, pigments, It may contain an odorant and a dye.

Another aspect of the present invention for achieving the above object is (a) extracting reeds with hot water; (b) cooling the extract; And (c) it provides a method for producing a cosmetic composition for skin whitening comprising the step of obtaining the cooled extract as a powder extract.

Specifically, the manufacturing method in the step (a), the high temperature may be 110 ℃ to 130 ℃, pressurization may be 10 psi to 20 psi, extracting for 10 minutes to 20 minutes under high temperature / pressure conditions In the step (b), the cooling may be performed at 0°C to ?80°C, and in the step (a), the obtained may be obtained through freeze-drying, but is not particularly limited thereto.

At this time, the terms “reed”, “high temperature”, “pressurized”, “extract”, “skin whitening” and “cosmetic composition” are as described above.

Another aspect of the present invention for achieving the above object provides a skin whitening method comprising administering a composition comprising the high temperature and pressurized hot water extract of reed to an individual other than a human.

At this time, the terms “reed”, “high temperature”, “pressurized”, “hot water”, “extract”, “skin whitening” and “cosmetic composition” are as described above.

In the present invention, "administration" means introducing the composition of the present invention to an individual by any suitable method, and the route of administration of the composition is to be administered through any general route as long as it reaches the target tissue and can exhibit skin whitening effect. I can. The administration may include application to the skin, and intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, or intranasal administration, but is not limited thereto. Specifically, due to the nature of the composition of the present invention having a skin whitening effect, the route of administration of the composition may be applied to the skin and administered.

The composition of the present invention may be administered daily or intermittently, and the number of administrations per day may be administered once or divided into 2 to 3 times. In addition, the composition of the present invention may be used alone or in combination with other substances for skin whitening. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.

In the present invention, "individual" refers to all animals such as rats, mice, livestock, including humans. Specifically, it may be a mammal including or not including humans.

Another aspect of the present invention for achieving the above object is to provide a quasi-drug composition for skin whitening, comprising the reed hot and pressurized hot water extract as an active ingredient.

At this time, the terms “reed”, “high temperature”, “pressurized”, “hot water”, “extract” and “skin whitening” are as described above.

In the present invention, the term "quasi-drug" refers to items that are less effective than pharmaceuticals among items used for the purpose of diagnosing, treating, improving, alleviating, treating or preventing diseases of humans or animals. For example, according to the Korean Pharmacological Law Quasi-drugs exclude items used for pharmaceutical purposes, and include products used for the treatment or prevention of diseases of humans and animals, and products with minor or no direct action on the human body.

The quasi-drug composition of the present invention may be prepared in a form selected from the group consisting of a body cleanser, foam, soap, mask, ointment, cream, lotion, essence, and spray, but is not limited thereto.

In the present invention of the present invention, when the composition containing the reed hot/pressed hot water extract as an active ingredient is used as a quasi-drug additive, the composition containing the reed hot/pressed hot water extract of the present invention as an active ingredient is added as it is or other quasi-drug or quasi-drugs It can be used together with an ingredient, and can be used suitably according to a conventional method.

Another aspect of the present invention for achieving the above object provides a pharmaceutical composition for the prevention or treatment of skin whitening-related diseases, comprising the reed hot and pressurized hot water extract as an active ingredient.

In the present invention, the term "whitening-related disease" refers to a disease having symptoms in which pigment is excessively deposited on the skin, and specifically, the whitening-related disease may be spots, freckles, age spots, blemishes, or melanoma.

In the present invention, the term "prevention" means any action in which the composition of the present invention inhibits or delays the occurrence of whitening-related diseases.

In the present invention, the term "treatment" refers to any action that makes the pharmaceutical composition of the present invention ameliorate or benefit the symptoms of whitening-related diseases.

In the present invention, the pharmaceutical composition may further include a pharmaceutically acceptable carrier.

In the present invention, the term "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not irritate an organism and does not inhibit the prophylactic or therapeutic activity and properties of whitening-related disease symptoms of the present composition. As acceptable pharmaceutical carriers for compositions formulated as liquid solutions, sterilization and biocompatible ones, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol And one or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as needed.

In addition, the pharmaceutically acceptable carrier of the present invention may include a non-natural carrier.

The pharmaceutical composition of the present invention may be administered singly or in multiple doses in a pharmaceutically effective amount.

In the present invention, the term "pharmaceutically effective amount" refers to an amount sufficient to prevent or treat a disease at a reasonable benefit/risk ratio applicable to medical prevention or treatment, and the effective dose level is the severity of the disease, the activity of the drug , Including the patient's weight, health, sex, sensitivity to the patient's drugs, the time of administration of the composition of the present invention used, the route of administration and the rate of excretion, the duration of treatment, and drugs used in combination or concurrent with the composition of the present invention used Factors and other factors well known in the medical field can be determined.

The composition of the present invention comprises a reed root extract extracted through high temperature and pressure as an active ingredient, and the extract has antioxidant effects, inhibits tyrosinase and α-glucosidase activities, and inhibits the activity of melanocytes It can be used for skin whitening to prevent and improve pigmentation.

1 is a process chart showing a process method of a high temperature and pressurized hot water extraction method (121°C).
2 is a graph comparing the DPPH scavenging ability according to the extraction method of reed extract.
Figure 3 is a graph comparing the hydrogen peroxide scavenging ability according to the extraction method of reed extract.
4 is a graph comparing the reducing power reducing power according to the extraction method of reed extract.
5 is a graph showing the evaluation of melanogenesis cytotoxicity of the reed ethanol (70%) extract.
Figure 6 is a graph showing the melanogenesis cytotoxicity evaluation of the reed ethanol (100%) extract.
7 is a graph showing the toxicity evaluation of melanogenic cells of hot water extract of reeds.
8 is a graph showing the melanin pigment inhibitory ability of the reed ethanol (70%) extract.
9 is a graph showing the melanin pigment inhibitory ability of the reed ethanol (100%) extract.
10 is a graph showing the melanin pigment inhibitory ability of reed hot water extract.
11 is a graph showing a comparison of in vitro tyrosinase activity inhibitory effects according to the extraction method of reed extract.
12 is a graph showing the inhibitory effect of α-glucosidase activity of hot water extract of reeds.
13 is a graph showing the cell tyrosinase inhibitory effect of B16F10 melanocytes in hot water extract of reeds.
14 is a graph showing changes in pH and specific gravity according to harsh and accelerated conditions of a formulation containing a reed root extract.
15 is a photograph showing the safety of the serum test formulation containing the reed root extract against general bacteria.
16 is a photograph showing the safety of the serum test formulation containing reed root extract against fungi.
17 is a graph comparing the coumarin acid content of the hot and pressurized hot water extract and the general hot water extract through HPLC analysis chromatogram.
18 is a graph comparing the L-DOPA oxidation inhibitory ability of hot and pressurized hot water extracts and general hot water extracts.

Hereinafter, the present invention will be described in more detail through the following examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example 1: Preparation of reed hot/pressurized hot water extract

Hot water extract, ethanol (70%) extract, and ethanol (100%) extract were prepared using reed root.

Specifically, the hot water extract was extracted using purified water at 121°C, and the ethanol extract was prepared by extraction at room temperature and pressure using 70% and 100% ethanol.

More specifically, the method of preparing a hot water extract was extracted by applying 10 kg of dried reed root and 200 L of purified water at 121°C and 15psi for 15 minutes, and the extract was filtered and cooled at -80°C, followed by lyophilization (Fig. 1). ).

Example 2: Comparison of antioxidant effects according to extraction methods

Example 2-1: Comparison of total phenol and total flavonoid contents according to the extraction method

Total phenol and total flavonoid contents according to the reed extract extraction method were compared.

More specifically, the total polyphenol content was analyzed according to the Folin-Denis method using the principle that a phenolic substance reacts with phosphomolybdic acid to develop a blue color. 0.5 mL of Folin reagent was mixed with 0.5 mL of the extract (1 mg/mL), reacted at room temperature for 3 minutes, 1.5 mL of 2% Na3CO3 was added, reacted in the dark for 2 hours, and then used a microplate reader at 760 nm. Then, the absorbance (OD) was measured. The total polyphenol content in the sample was expressed as gallic acid equivalent (GAE) per gram of sample through the gallic acid standard curve.

In addition, for the analysis of the total flavonoid content, 1 mL of 2% aluminum chloride methanolic solution was mixed with 1 mL of the extract, followed by reacting at room temperature for 15 minutes, and absorbance was measured at 430 nm using a microplate reader (Tecan Group Ltd.). The total flavonoid content in the sample was expressed as quercetin equivalent (QE) per gram of sample through the standard curve of quercetin.

As a result, the hot water extract of reeds was 387.3 μg GAE/mg of total phenol and 12.56 μg GAE/mg of total flavonoids, confirming the highest content in the extract.

Through this, it was predicted that the content of the effective physiologically active substance was the highest in the hot water extract of reeds, and the antioxidant effect would be highest.

Example 2-2: DPPH removal rate measurement

DPPH scavenging rates according to the reed extract extraction method were compared.

Specifically, the total DPPH scavenging rate was analyzed by comparing the antioxidant efficacy with an SC ₅₀ value. More specifically, the SC ₅₀ was reacted in the dark for 30 minutes after adding a DPPH (2,2-diphenyl-1-picrylhydrazyl) solution to the sample on a 96-well micro plate, and mixing well. After measuring the absorbance at 570 nm, the reaction solution was expressed as radical scavenging activity (%) and 50% scavenging concentration (SC ₅₀ ).

As a result, also as shown in Figure 2, the reed ethanol (70%), extract of the SC ₅₀ is 0.755 mg / mL, reed ethanol (100%), extract of the SC ₅₀ is 0.955 mg / mL, the reed hot water extract SC ₅₀ is 0.707 It was confirmed that it was mg/mL.

Through this, it was confirmed that the highest DPPH scavenging ability appeared in the hot water extract of reeds.

Example 2-3: Hydrgogen peroxide scavenging ability measurement

The hydrogen peroxide scavenging ability according to the extraction method of reed extract was compared.

Specifically, the total hydrogen peroxide scavenging ability was analyzed by comparing the antioxidant efficacy with an SC ₅₀ value. SC ₅₀ is as described above.

As a result, as can be seen in Figure 3, the IC50 of the reed ethanol (70%) extract is 1.004 mg / mL, the SC ₅₀ of the reed ethanol (100%) extract is 0.973 mg / mL, the SC ₅₀ of the hot water extract of reed is 0.723 mg It was confirmed that it was /mL.

Through this, it was confirmed that the highest hydrogen peroxide scavenging ability appeared in the hot water extract of reeds.

Example 2-4: Reducing power measurement of reducing power

Reducing power reduction power according to the reed extract extraction method was compared.

Specifically, the reducing power was measured through the hydrogen peroxidase scavenging method, and more specifically, after reacting for 5 minutes by adding 20% hydrogen peroxide solution to the sample on a 96-well micro plate, ABTS and peroxidase were added to react at 37 for 10 minutes. Then, the reaction solution was measured for absorbance at 405 nm.

As a result, as shown in Figure 4, it was confirmed that the absorbance of the reed ethanol (70%) extract at 200 μg/mL concentration was 0.199, the absorbance of the reed ethanol (100%) extract 0.186, and the reed hot water extract 0.213 .

Through this, it was confirmed that the highest reducing power appeared in the hot water extract of reeds.

Example 3: Cytotoxicity evaluation according to the extraction method

Cytotoxicity was compared according to the extraction method of reed extract.

More specifically, in order to compare the cytotoxicity according to the reed extract extraction method, the cytotoxicity was compared by treating the reed extract on B16F10 melanogenic cells.

As a result, as can be seen in FIGS. 5 to 7, when 400 ug/mL of reed ethanol (70%) extract was treated for 24 hours, when treated for 29% and 48 hours, 50% cytotoxicity was observed, and reed ethanol (100%) When 400 ug/mL of extract was treated for 24 hours, when 33% and 48 hours were treated, cytotoxicity of 60% was observed, and when 400 ug/mL of reed hot water extract was treated for 24 hours, 19% , When treated for 48 hours, it was confirmed that 50% of cytotoxicity appeared.

Through this, it was confirmed that the hot water extract is the safest in the cytotoxicity evaluation of melanocytes according to the three extraction methods.

Example 　 4: whitening effect according to the extraction method

Example 4-1: Comparison of melanin pigment generation amount evaluation

The ability to inhibit melanin pigment according to the reed extract extraction method was compared.

More specifically, in order to compare the melanin inhibitory ability according to the reed extract extraction method, B16F10 melanin-forming cells were treated with α-MSH to induce melanin production, and the reed extract was treated to compare the melanin inhibitory ability. At this time, arbutin was used as a positive control, and the comparison was performed using a standard curve of melanin pigment.

As a result, as can be seen in FIG. 8, the reed ethanol (70%) extract significantly decreased compared to α-MSH at 25 and 50 ug/mL concentrations, but at a concentration of 100 ug/mL or higher, the melanin production inhibitory effect was significantly reduced. It was confirmed that it did not appear.

In addition, as can be seen in FIG. 9, it was confirmed that the reed ethanol (100%) extract showed an inhibitory control effect of approximately 10%, compared to α-MSH, and had no significance.

In addition, as can be seen in Figure 10, the reed hot water extract showed significant melanin inhibitory ability at all concentrations of 25, 50, 100 and 200 ug/mL, and produced melanin similar to arbutin at concentrations of 25, 50 and 100 ug/mL. It was confirmed that at a concentration of 200 ug/mL, melanogenesis was reduced by more than 3% than arbutin.

Through this, it was confirmed that the reed hot water extract has superior melanin production reduction effect than the reed ethanol extract (70, 100%).

Example 4-2: Comparison of inhibition of tyrosinase activity

The inhibitory efficacy of tyrosinase activity according to the reed extract extraction method was compared.

Specifically, based on the KFDA data, the inhibitory effect of tyrosinase activity was compared through IC50 through an in vitro experiment.

As a result, as can be seen in Figure 11, it was confirmed that the SC ₅₀ of the reed hot water extract is 0.533 mg/mL.

Through this, it was confirmed that the reed hot water extract has the highest tyrosinase activity inhibitory effect among the reed extracts, and the tyrosinase activity inhibitory effect increases depending on the concentration.

Example 5: α-glucosidase activity inhibition efficacy

The inhibitory effect of α-glucosidase activity of the hot water extract of reeds was confirmed.

Tyrosinase is produced by the N-linked glycosylation reaction process, at this time, α-glucosidase breaks the bond between mannose and glucose, thereby starting the N-linked glycosylation reaction, and α -It is known that inhibition of the activity of the glucosidase enzyme inhibits the glycosylation reaction of tyrosinase, and the inactivated form of tyrosinase moves to the melanosomes, thereby inhibiting pigmentation. Thus, the α-glucosidase activity inhibition effect was confirmed through SC ₅₀ , and Qucetin was used as a positive control.

As a result, as shown in FIG. 12, it was confirmed that α-glucosidase SC ₅₀ of the hot water extract of reed was 0.47 mg/mL, and α- of 12.33% at 100 ug/mL concentration and 22.71% at 200 ug/mL concentration. It was confirmed that there is a glucosidase inhibitory effect.

Through this, it was confirmed that the reed hot water extract has a whitening effect by inhibiting α-glucosidase in a concentration-dependent manner.

Example 6: Efficacy of Inhibiting Cellular Tyrosinase of Hot Water Extract

The tyrosinase inhibitory effect of the reed hot water extract was confirmed.

Specifically, melanocytes B16F10 were treated with the reed hot water extract of the present invention to compare the relative activity of tyrosinase in cells. More specifically, α-MSH was treated to induce melanin formation, and arbutin was used as a positive control.

As a result, as can be seen in FIG. 13, significance was observed at 100 and 200 ug/mL concentrations of the reed hot water extract, and tyrosinase inhibitory efficacy similar to arbutin in the positive control was observed at 100 ug/mL concentration, and 200 ug/mL In mL, it was confirmed that the tyrosinase inhibitory effect increased by more than 15% than arbutin.

Through this, it was confirmed that the reed hot water extract has the effect of inhibiting the tyrosinase activity increased by α-MSH, and it was confirmed that there is a whitening effect by reducing melanin production.

Example 7: Test formulation preparation and stability investigation

A test formulation containing the reed root extract of the present invention was prepared (Table 2), and a stability test was performed to investigate the stability thereof.

More specifically, a skin patch irritation stability investigation was requested to the Oriental Medicine Bio Industry Clinical Support Center, and the harsh/accelerated stability and formulation/indicator stability were tested according to the safety test guidelines of the Ministry of Food and Drug Safety (KFDA). The microbial safety of the serum test formulation containing the root extract was tested.

As a result, it was confirmed that it is safe without irritation by investigating the stability of skin patch irritation at the Clinical Support Center for Oriental Medicine Bio Industry.

In addition, as can be seen in FIG. 14, it was confirmed that the reed root extract did not affect the formulation without changes in pH and specific gravity under accelerated/severe conditions according to the safety test guidelines of the Ministry of Food and Drug Safety.

In addition, as can be seen in Figures 15 to 16, it was confirmed the stability of the serum test formulation containing the reed root extract against common bacteria and fungal microorganisms.

Through this, it was confirmed the stability of the test formulation containing the reed root extract of the present invention.

Example 8: Comparison of general hot water extraction method and high temperature/pressure hot water extraction method

Example 8-1: Comparison of coumarin acid content

In order to confirm the superiority of the hot/pressurized hot water extraction method, the extract through the hot/pressurized hot water extraction method of Example 1 and the coumarin acid of the extract extracted through the general hot water extraction method extracted at 100° C. without applying pressure were compared.

More specifically, the coumarin acid content was confirmed through HPLC analysis. The powder extract obtained through each extraction method was dissolved in ethanol at a concentration of 0.1% and analyzed by HPLC analysis.

As a result, as can be seen in Tables 3 to 17, compared to the general hot water extraction method, compared with the general hot water extraction method showing the coumarin acid content per 1 kg of lyophilisate is 2741 mg / kg 553 mg / kg in the high temperature / pressurized hot water extraction method It was confirmed that it was superior to 5 times or more, and it was confirmed that the hot/pressurized hot water extract was converted into a powder form after freeze-drying, whereas the general hot water extract was confirmed that the powder obtained after freeze-drying turned into a hard mass.

Through this, it was confirmed that the coumarin acid content of the hot/pressurized hot water extract was 5 times higher, and it was converted into a stable powder form, so that it was easy to use.

Example 8-2: Comparison of L-DOPA oxidation assay

In order to confirm the excellence of the hot/pressurized hot water extraction method, the L-DOPA oxidation assay of the extract through the hot/pressurized hot water extraction method of Example 1 and the extract extracted through the general hot water extraction method extracted at 100°C without applying pressure was compared. .

More specifically, samples obtained through the above method were prepared by diluting each of them in 0.1M sodium phosphate buffer (pH7.0) from 1000 μg/ml to a constant multiple, and tyrosinase, an enzyme using L-DOPA as a substrate, was 268.7mU solution, The substrate L-DOPA was prepared in 10 mM. In addition, 900 μg/mL of arbutin was prepared as a positive control. Thereafter, 5 μL of a tyrosinase solution was added to a 96-well plate, a sample or a positive control was added, and then 5 μL of L-DOPA and 85 μL of buffer were simultaneously added. As a blank test solution, buffer was added instead of L-DOPA for analysis. Then, the reaction was performed at room temperature for 20 to 30 minutes, and the absorbance value was measured at 450 nm with an ELISA reader. L-DOPA oxidation (%) was calculated through the following formula.

L-DOPA oxidation (%) = {(test group-blank test group of test group)/(negative control group-blank test group of negative control group) *100}

As a result, as can be seen in FIG. 18, the hot/pressurized hot water extract showed inhibitory activity from 200 μg/mL, but the general hot water extract showed inhibitory activity from 500 μg/mL, and the hot/pressed hot water extract was general hot water extract. It was confirmed that higher inhibitory activity appeared.

Through this, it was confirmed that the high-temperature/pressurized hot-water extract showed higher antioxidant activity than the general hot-water extract.

From the above description, those skilled in the art to which the present invention pertains will be able to understand that the present invention can be implemented in other specific forms without changing the technical spirit or essential features thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not limiting. The scope of the present invention should be construed that all changes or modifications derived from the meaning and scope of the claims to be described later rather than the above detailed description and equivalent concepts are included in the scope of the present invention.

Claims (19)

A cosmetic composition for skin whitening that contains hot water extract of the reed (Phragmites Communis) root as an active ingredient,
The hot water extract is to extract hot water for 10 to 20 minutes at a high temperature of 110°C to 130°C and pressurized conditions of 15 psi to 20 psi,
The cosmetic composition, characterized in that the coumarin acid content is increased.
delete delete delete delete The cosmetic composition of claim 1, wherein the extract is in powder form.
The cosmetic composition of claim 6, wherein the powder is obtained through lyophilization.
The cosmetic composition of claim 1, wherein the extract has an antioxidant effect.
The cosmetic composition of claim 1, wherein the whitening is due to inhibition of the activity of tyrosinase.
The cosmetic composition according to claim 1, wherein the whitening is due to inhibition of the activity of α-glucosidase.
The cosmetic composition of claim 1, wherein the whitening is due to inhibition of the activity of melanocytes.
delete (a) extracting hot water from the reed (Phragmites Communis) roots at a high temperature of 110°C to 130°C and pressurization conditions of 15psi to 20psi for 10 to 20 minutes; (b) cooling the extract; And (c) obtaining the cooled extract as an extract in powder form,
The obtained extract is characterized in that the content of coumarin acid is increased, the method for producing a cosmetic composition for skin whitening.
delete delete delete The method of claim 13, wherein the cooling in step (b) is performed at 0°C to 80°C.
The method of claim 13, wherein the step (c) is obtained through lyophilization.
It is a quasi-drug composition for skin whitening that contains the hot and pressurized hot water extract of the root of Phragmites Communis as an active ingredient,
The hot water extract is to extract hot water for 10 to 20 minutes at a high temperature of 110°C to 130°C and pressurized conditions of 15 psi to 20 psi,
It is characterized in that the coumarin acid content is increased, quasi-drug composition.

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