KR20170001649A

KR20170001649A - Composition comprising herbal extract for improving skin wrinkle and moisturizing

Info

Publication number: KR20170001649A
Application number: KR1020160079647A
Authority: KR
Inventors: 이정복; 하헌용
Original assignee: 이정복
Priority date: 2015-06-25
Filing date: 2016-06-24
Publication date: 2017-01-04

Abstract

The present invention relates to a composition for improving and moisturizing wrinkles containing a mixed herbal extract, and more particularly to a composition for improving and moisturizing wrinkles containing ginseng extracts, ginseng extracts, ginseng extracts, ginseng extracts, The present invention relates to a cosmetic composition for improving and moisturizing or a composition for external application for skin. The herbal mixed extract according to the present invention has advantages of improving skin wrinkles, moisturizing effect, skin toxicity, and less side effects as a natural product.

Description

[0001] The present invention relates to a composition for improving wrinkles and moisturizing,

The present invention relates to a composition for improving and moisturizing wrinkles containing a mixed herbal extract, and more particularly to a composition for improving and moisturizing wrinkles containing ginseng extracts, ginseng extracts, ginseng extracts, ginseng extracts, Improving and moisturizing cosmetic composition or external composition for skin.

Skin is a body wrapped around the body, directly in contact with the external environment while protecting the body from various harmful factors is responsible. The skin system is an organs covering the outside, consisting of three distinct layers of skin, dermis and subcutaneous fat layer from the outside. The epidermis is mostly keratinized cells of middle layer squamous epithelium. The dermis, composed of matrix proteins such as collagen fibers and elastic fibers, is located beneath the epidermis, and the dermis contains blood vessels, nerves, and sweat glands. The subcutaneous fat layer is composed of adipocytes. The skin has various functions such as controlling the body temperature and functioning as a barrier against the external environment.

However, skin is increasingly damaged by age, ultraviolet rays, external pollutants and stress, and the function of protecting the skin from these factors is weakened, thereby promoting the skin aging phenomenon as the cell protection and proliferation ability deteriorates. This aging phenomenon is mainly due to the hyaluronic acid cleavage and low molecular weight and the collagen production of the fibroblasts, resulting in a decrease in the water retention capacity of the keratinocyte and dermis layers, The skin elasticity is reduced by weakening.

Generally, the moisture content of the skin is about 70% in the dermal layer, but decreases to the skin layer, which is about 10% to 30% in the keratin layer. The water supplied from the dermis layer mainly migrates to the upper part of the stratum corneum by passive diffusion and eventually to the outside. This transdermal water loss (TEWL) is known to be maintained at an appropriate level by lipid component of epidermal keratinocyte and epidermal lipid. In addition, a substance having a hydrophilic water retention capacity called a Natural Moisturizing Factor (NMF) is present in the keratinocyte layer, and it is known that it plays an important role in moisturizing the skin.

When the moisture of the keratin layer is maintained at about 10% to 30%, the skin is smooth and soft, and the body protection functions normally. However, when the moisture content of keratinocytes is less than 10%, the skin becomes rough and loses protective function of the body and aging occurs. For example, in the case of dry skin, the keratinocyte aggregation ability is weakened and a scale phenomenon appears in which the keratinocyte is peeled off like a piece of scales from the surface of the skin. The dryness of the skin in this way indicates that the moisture content of keratinocyte layer It is less than normal skin. In addition to this, even healthy skin may be in a harsh environment, such as wind, cold weather, sunshine, cleansing, shaving, etc., which causes water shortage, which can cause a bad skin condition. Therefore, it is very important to keep the moisture content of keratinocytes in skin properly.

To this end, cosmetics were added with ingredients similar to sebum, NMF ingredients, or moisturizers such as polyols. For example, glycerin, sorbitol and the like having three or more hydroxyl groups (OH groups) as the water-soluble polyol are added. They show excellent moisture retention, but they are very sticky and have a problem of feeling uncomfortable when used and depriving the skin of moisture , And propylene glycol having a hydroxyl group (OH group) of 2, 1,3-butylene glycol and the like may cause side effects of the skin. In addition, typical wrinkle prevention and improvement cosmetics include vitamin A (retinol) or retinyl palmitate. However, the products containing these raw materials are not only sensitive to changes over time, but also have a limited amount of use due to problems of stability such as skin irritation and redness when they are penetrated or decomposed into the skin at an effective concentration or more, And the effect of moisturizing ability was minimal. In addition, additives such as sodium pyrrolidonecarboxylate (PCA-Na), sodium lactate, and urea, which are other natural moisturizing factors, have a problem of deteriorating the emulsification stability of cosmetics due to their high electrolytic properties. Amino acids, collagen, elastin Even though it has a moisturizing ability, its moisturizing ability is limited.

Recently, the development of a cosmetic for preventing skin aging has been actively developed by introducing a physiologically active substance obtained from plants, microorganisms, animals, etc. into cosmetics to maintain the inherent defensive function of the skin itself, thereby enhancing the moisturizing power and elasticity.

Korean Patent Publication No. 2008-0036694 (Published April 29, 2008) Korea Registered Patent No. 1,435,679 (registered on August 22, 2014) Korean Registered Patent No. 889,605 (Registered on March 12, 2009) Korea Registered Patent No. 1,456,052 (Registered on October 23, 2014) Korean Patent Publication No. 2009-0130584 (disclosed on December 24, 2009)

An object of the present invention is to provide a cosmetic composition and a composition for external application for skin excellent in the improvement of skin wrinkles and moisturizing effect.

Another object of the present invention is to provide a cosmetic composition for skin wrinkle improvement and moisturizing and a composition for external application for skin derived from natural materials without cytotoxicity and without side effects.

The present invention has solved the above problems by providing a cosmetic composition for improving and moisturizing wrinkles and a composition for external application for skin containing a mixed extract of ginseng root, ginseng root, peony root, Angelica gigas Nakai, bamboo root, dermis,

The mixed extract of herbal medicine according to the present invention is advantageous in improving skin wrinkles, moisturizing effect, no skin toxicity, and having few side effects as a natural product.

FIG. 1 shows the results of MTT analysis of licorice, Hwanggi, and herbal medicine mixed extract according to the present invention.
Fig. 2A shows the result of hyaluronic acid ELISA of licorice root and licorice root.
2B shows the results of measurement of the degree of synthesis of hyaluronic acid in Angelica keiskei koidz.
Fig. 3 shows the result of measurement of hyaluronidase inhibition rate of licorice.
FIG. 4 and FIG. 5 show experimental results of measuring the HAS2 gene expression pattern.

The present invention relates to a cosmetic composition for improving and moisturizing wrinkles containing a mixed extract of ginseng, hwanggi, peony root, Angelica gigas Nakai, Baekjong, dyke, dermis, allium, gilt eum, white and licorice.

In one embodiment of the present invention, ginseng, hwanggi, peony, angelica, chrysanthemum, dwarf, dermis, allium, gilt eum, white paper and licorice can be extracted with water, C ₁ _-2 lower alcohol or a mixed solvent thereof. The C ₁ _-2 lower alcohol is methanol or ethanol, more specifically, ethanol can be used.

In one embodiment of the present invention, the content of ginseng, hwanggi, peonies, angelica, gypsum, bamboo, dandelion, allium, gilt silver, white paper and licorice is from 1.5 to 9 parts by weight: 3 to 15 parts by weight: 3 to 10 parts by weight: 3 to 15 parts by weight: 3 to 15 parts by weight: 3 to 15 parts by weight: 3 to 9 parts by weight: 3 to 18 parts by weight: 3 to 9 parts by weight: 3 to 10 parts by weight .

In one embodiment of the present invention, the amount of the extracting solvent is preferably 5 to 15 times the total dry weight of the ginseng, the herb, the peony, the ginseng, the ginseng, the dandelion, the tussock, the ginseng, It is more preferable to add 10 times the dry weight, but it is not limited thereto.

The temperature for extraction is preferably 10 to 120 ° C, but is not limited thereto, and is more preferably 40 to 100 ° C, but is not limited thereto. The extraction time is preferably 1 hour to 7 hours, more preferably 3 hours to 5 hours, but is not limited thereto, and most preferably 3 hours, but is not limited thereto.

The method for producing the extract may be a method using an extraction device such as hot water extraction, immersion extraction, reflux cooling extraction, supercritical extraction, subcritical extraction, high temperature extraction, high pressure extraction or ultrasonic extraction, or adsorption resin including XAD and HP- And a method using a conventional extraction method such as a method using the above-mentioned method.

In one embodiment of the present invention, the composition exhibits an activity of inhibiting hyaluronidase activity.

As a specific formulation of the cosmetic composition of the present invention, there may be mentioned a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, And includes formulations such as soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, milky lotion, press powder, loose powder, eye shadow and the like.

The cosmetic composition of the present invention may further comprise at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickeners and gelling agents as well as mixed extracts of ginseng, hwanggi, peony, angelica, A stabilizer, a foaming agent, a perfume, a surfactant, water, an ionic or nonionic emulsifier, a filler, a sequestering and chelating agent, a preservative, a vitamin, a barrier agent, a humectant, an antioxidant, a suspending agent, , Essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients commonly used in cosmetics.

The present invention relates to a composition for external application for skin improving and moisturizing, which comprises a mixed extract of ginseng, hwanggi, peony root, Angelica gigas, Baekyoung, dongryeong, dermis, allium, gilt eum, white and licorice.

When the mixed extract of ginseng, hwanggi, peony, angelica, lepidoptera, bepyeong, dandelion, allium, gilt eum, white and licorice of the present invention is used as an external preparation for skin, it may further contain a fatty substance, an organic solvent, a solubilizing agent, , Emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, May contain adjuvants conventionally used in the field of dermatology, such as topical agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredient commonly used in external preparations for skin. In addition, the components can be introduced in amounts commonly used in the dermatology field.

Hereinafter, the present invention will be described in more detail with reference to examples. The following examples are intended to aid the understanding of the present invention and are not intended to limit the scope of the present invention thereto.

Example 1 . mix Herbal medicine Preparation of composition

<1-1> 70% ethanol extract

The ginseng, Huangji, peonies, angelica, angelica, gingiva, dandelion, allium, gingkohwa, white ginseng and licorice contained in the mixed extract of the present invention were selected and controlled according to the degree of pharmacopoeia. Each herbal medicine was added to the extractor in an amount of 10 g of 70% (by weight) of the drug, and the amount of the herbal medicine was adjusted to 10 g of the drug by adding 125 g of peony root, 125 g of Angelica gigasum, 125 g of alfalfa, 125 g of dandruff, 125 g of alabaster, 125 g of Ganoderma lucidum, v / v) ethanol (12.5 L) was added thereto, followed by extraction at 80 ° C for 3 hours, followed by concentration and drying. The dry extracts were dissolved in concentrations of 70% ethanol just prior to in vivo and in vivo experiments. Hereinafter, the experiment of the examples was carried out using a 70% ethanol extract.

&Lt; 1-2 > 70% methanol extract

The ginseng, Huangji, peonies, angelica, angelica, gingiva, dandelion, allium, gingkohwa, white ginseng and licorice contained in the mixed extract of the present invention were selected and controlled according to the degree of pharmacopoeia. Each herbal medicine was added to the extractor in an amount of 10 g of 70% (by weight) of the drug, and the amount of the herbal medicine was adjusted to 10 g of the drug by adding 125 g of peony root, 125 g of Angelica gigasum, 125 g of alfalfa, 125 g of dandruff, 125 g of alabaster, 125 g of Ganoderma lucidum, v / v) methanol (12.5 L) was added and sonicated at 40 ° C for 3 hours. The extract was filtered, and the filtrate was concentrated under reduced pressure at 60 ° C or lower.

<1-3> Hot water extract

The ginseng, Huangji, peonies, angelica, angelica, gingiva, dandelion, allium, gingkohwa, white ginseng and licorice contained in the mixed extract of the present invention were selected and controlled according to the degree of pharmacopoeia. 125 g of peony root, 125 g of Angelica gigasum, 125 g of alfalfa, 125 g of dandelion, 125 g of brassiere, 125 g of ginger silver, 63 g of white paper and 63 g of licorice were put into the extractor and purified water of 12.5 L), and the mixture was extracted at 100 ° C for 3 hours. The extract was filtered, and the filtrate was concentrated under reduced pressure at 60 ° C or lower.

Experimental Example 1. Basic functional evaluation test for herbal materials

<1-1> Preparation of herbal medicine extract

100 g of medicinal herbs were measured, leached out as a solvent, filtered and collected in a bottle, and this process was repeated three times. After concentration with a vacuum concentrator, it was lyophilized. The yield of the final extract was calculated by measuring the mass. More specifically, the ginseng, Huangji, peony, angelica, lily, bamboo, dandelion, allium, gold, silver, white and licorice contained in the mixed extract of the present invention are selected, 200 g of each of phytophthora, angelica, angelica, lycopene, dandelion, allium, gold, silver, white and licorice were separately placed in an extractor and immersed in 70% (v / v) ethanol in 10 times the weight of the drug and extracted at 80 ° C for 3 hours Followed by concentration and drying. The dry extracts were dissolved in concentrations of 70% ethanol just prior to in vivo and in vivo experiments.

Extraction yield (%) according to the manufacturing method of single component extract Ginseng Hwanggi Peony Angelica Boiled dermis Abalone Gold silver blank licorice yield 26 12 25.7 33 10 22 23 25 38.5 20

<1-2> Culture of HaCaT cells

HaCaT cells were cultured in Dulbecco's Modified Eagle Medium (DMEM, Gibco) 10% Fetal bovine serum (FBS, Gibco) and 1% penicillin-streptomycin (PS, Gibco) with 37 ℃ in a culture medium, 5% CO ₂ condition contained in .

<1-3> MTT analysis

Cells were seeded into 96 well plates at a concentration of 2 x 10 < ⁵ > / mL. After 24 hours, the cells were replaced with serum-free DMEM. For 24 hours to give a kick out the culture medium after haejun put 3- (4,5-dimethyl-2- thiazolyl) -2,5-diphenyl-2H-tetrazolium bromide (MTT, 5 mg / ml) in 20 μl CO ₂ And cultured in an incubator for 2 hours. After dissolving the crystals in 100 μl of DMSO, the absorbance was measured at 570 nm. The DMSO contained in the sample treated with the cells was adjusted to a final concentration of 0.1%.

<1-4> ELISA

Cells were seeded into 6 well plates at a concentration of 2 x 10 < ⁵ > / mL. After 24 hours, the cells were washed twice with serum-free DMEM and then replaced with serum-free DMEM and the samples were treated. DMSO was adjusted to 1%. After 24 hours, 350 μL of medium was removed, and the same volume was removed 24 hours later. Centrifugation at 15,000 xg for 5 min. The supernatant was removed and stored at -20 ° C until ELISA. The ELISA was performed using the HA-ELISA kit (echelon) and the manufacturer provided method. ATRA 1 M was used as a positive control.

<1-5> RT-PCR

Cells were seeded into 6 well plates at a concentration of 2 x 10 < ⁵ > / mL. After 24 hours, the cells were replaced with serum-free DMEM. Final DMSO was adjusted to a concentration of 0.1%. After incubation for 24 hours, RNA was extracted with easyBlue (intron). After measuring the RNA concentration and purity (OD ₂₆₀ / OD ₂₈₀ ), cDNA was synthesized with 2 μg RNA using a power cDNA synthesis kit (intron). PCR was performed using a premix PCR kit (Solgent). The PCR product was electrophoresed on 1.5% agarose gel to confirm the band. All-trans-retinoic acid 1 M (ATRA, Sigma) was used as a positive control. Primer was constructed with the sequence reported in the previous paper.

Gene Direction Sequence (5 '- > 3') Size (bp) HAS2 Forward GCT ACC AGT TTA TCC AAA CG (20 mer) 393 Reverse GTG ACT CAT CTG TCT CAC CG (20 mer) GAPDH Forward ATT GTT GCC ATC AAT GAC CC (20 mer) 546 Reverse AGT AGA GGC AGG GAT GAT GT (20 mer)

The PCR conditions were as follows. 94 ° C 15 min, 32-35 cycles: 94 ° C 30s 50 ° C 30s 72 ° C 60s 72 ° C 10 min Final synthesis.

The extraction yields are as shown in the following table:

Extraction solvent
sample 80% EtOH 100% MeOH 80% MeOH Licorice-1 12.6% 16.5% 26.7% Licorice-2 13.8% 11.5% 24.0% Angelica -1 32.8% 30.9% 44.5% Angelica-2 36.7% 38.5% 49.1% Hwanggi-1 20.9% 18.5% 31.2% Hwanggi-2 12.0% 9.4% 17.8%

<1-6> Inhibition of hyaluronidase activity

The increase of HA content can be thought of as the increase of HA production and the decrease of degradation. HA reduction is known to be the action of HYAL. The inhibitory effects of licorice extracts were 49.1, 33.86 and 10.36% at the treatment concentrations of 500 μg / mL, 200 μg / mL and 100 μg / mL, respectively. At 100 μg / mL, the licorice extract inhibited HYAL by 10.36%. As shown in the above MTT results, the toxicity to HaCaT was very strong at this concentration.

<1-7> RT-PCR

To determine whether the HAS2 bands could be identified by RT-PCR, RT-PCR was performed after treatment with ATRA 0.1 μM and 1 μM ATRA, which was already known to induce HAS2 gene expression. The previously reported HAS2 primer (1) was used. 1 μL of cDNA prepared with 2 μg of RNA was subjected to PCR at 94 ° C for 15 min, 35 cycles: 94 ° C for 30 s, 50 ° C for 30 s, 72 ° C for 60 s, and 72 ° C for 10 min. Other bands on the gel were not visible to the naked eye and the predicted 393 bp HAS2 and 546 bp GAPDH bands were compared to the markers. Therefore, the primer and temperature conditions are considered to be appropriate, and as expected, the band is darker than the 0.1 μM treatment at ATRA 1 μM treatment. The final PCR conditions were 3 μL of cDNA prepared with 2 μg RNA, and the number of cycles was 32 at the same temperature condition.

The expression level of HAS2 gene was confirmed by RT-PCR when 50%, 100%, or 200 μg / mL of methanol extract of 80% methanol was treated. The expression of HAS2 was markedly increased when treated with ATRA 1 μM, a positive control, and the expression of HAS2 gene was increased when 200 μg / mL was treated within the treatment concentration range.

Experimental Example 2 . Moisturizing test

To evaluate the efficacy and safety of the cosmetic composition according to the present invention, a comparative test with a reference product was conducted in a randomized, single-blind, double-blind fashion. The test center is the RIC Clinical Trials Center of Semyung University, and the study period, test design, etc. are as shown in the following table:

Research period January 01, 2015 - July 31, 2015 Test design Single-institution, randomized, double-blind, comparative clinical trials Research Purpose To evaluate the efficacy and safety, such as improvement of skin health (moisturizing, elasticity, dermal density) when a cosmetic according to the present invention was used for 8 weeks in a female subject. Number of subjects The number of subjects who can be finally assessed is 20, 20% Subject Non-lactating women of 30 to 50 years of age who are not pregnant Subject selection criteria (Subject subjects should comply with all of the following criteria)
1. Age: Female under 30 to 50 years old
2. Subjects who use functional cosmetics for the purpose of improving skin wrinkles and skin elasticity, or who are not taking health functional foods or similar products for skin health
3. Healthy subjects without skin disorders, including skin diseases
4. A person who has decided to participate in the examination after hearing the detailed explanation of the human body test and has signed the consent form Exclusion criteria for subjects (Subjects with any of the following can not participate in this study.)
1. Patients with a history of drug hypersensitivity
2. Subjects who have or have undergone hormone replacement therapy within 3 months
3. Subjects with heart disease (heart failure, angina pectoris, myocardial infarction)
4. Unadjusted hypertensive subjects (> SBP 145 mmHg or <DBP 90 mmHg)
5. Malignant tumors, subjects with narrow angle glaucoma or lung disease
6. Subjects with severe renal impairment or liver dysfunction (serum creatinine> 2.0 mg / dl, ALT, AST, alkaline phosphatase> normal upper limit of 2.5)
7. Subjects with neurological or psychiatric history or who are currently suffering from the disease (schizophrenia, epilepsy, alcoholism, drug addiction, anorexia, abnormal appetite stimulation, etc.)
8. Subjects with a history of stroke or transient ischemic heart attack
9. If you have been taking medications that affect the absorption, distribution, metabolism, or excretion of oral or injectable steroids, hormones, or drugs within the last 3 months, or have taken medications that could affect your skin, or participated in other human trials Experienced subjects
10. Subjects receiving other forbidden treatments (insulin, antidepressants, antispotagens, barbiturates, antipsychotics, drugs of abuse)
11. Subjects who have been using steroid-containing skins for more than one month to treat skin diseases
12. Subjects with sensitive, irritable skin
13. Subjects with skin abnormalities such as dots, acne, erythema and capillary dilatation on the test site
14. Subjects who received skin peeling or wrinkle removal procedures within 6 months before the start of the study
15. Prohibited pregnant women, lactating women, pregnant women, or those who do not consent to appropriate contraceptive methods (eg oral contraceptives, hormonal implants, intrauterine devices, condoms, spermicides)
16. According to the opinion of the person in charge of the human test, the person who is judged to be unable to comply with the test or who is deemed inappropriate by the doctor Clinical trial
product Test product: Product containing herbal medicine mixed extract according to the present invention
○ Control Products: Control Products Use period and
How to use ○ Test product: Apply to skin for 8 weeks 3 times a day.
○ Control product: Apply to the other side skin for 8 weeks 3 times a day.

evaluation Way Efficacy
① Primary efficacy evaluation (Primary endpoint)
○ Assess the skin moisture content of the test group and the control group after baseline and after 8 weeks.

② Secondary endpoint
Baseline and after 8 weeks test group and control group
○ Skin elasticity
○ Density of dermis
○ Question evaluation
And so on.

Safety
○ Before and after using the test product and the control product, after 8 weeks of use, perform an experimental test and a vital sign test and evaluate the adverse reaction throughout the study period

[Clinical Test Results]

Claims

A cosmetic composition for improving and moisturizing wrinkles containing a mixed extract of ginseng, hwanggi, peony root, Angelica gigas, Baekyoung, dyke, dermis, allium, gilt eum, white and licorice.

The method according to claim 1,
A cosmetic composition for improving and moisturizing wrinkles, characterized in that it is extracted with water, C ₁ _-2 lower alcohol or a mixed solvent thereof, which comprises ginseng, hwanggi, peony root, Angelica gigas, Baekyoung, .

The method according to claim 1,
1.5 to 9 parts by weight 3 to 15 parts by weight 3 to 10 parts by weight 3 to 24 parts by weight 3 to 15 parts by weight of ginseng Part 3 to 9 parts by weight 3 to 15 parts by weight 3 to 9 parts by weight 3 to 18 parts by weight 3 to 9 parts by weight 3 to 10 parts by weight wrinkle improving and moisturizing / RTI >

The method according to claim 1,
Wherein the composition inhibits hyaluronidase activity. &Lt; RTI ID = 0.0 > 11. < / RTI >

A composition for external application for skin for improving and moisturizing wrinkles containing a mixed extract of ginseng, hwanggi, peony root, Angelica gigas, Baekyoung, dynasty, dermis, allium, gilt eum, white and licorice.

6. The method of claim 5,
Wherein the composition is extracted with water, a C ₁ _-2 lower alcohol or a mixed solvent thereof, wherein the composition is selected from the group consisting of ginseng, hwanggi, peony root, Angelica gigas, Baekjeong, gyeongryeong, dermis, allium, Composition.

6. The method of claim 5,
1.5 to 9 parts by weight 3 to 15 parts by weight 3 to 10 parts by weight 3 to 24 parts by weight 3 to 15 parts by weight of ginseng Part 3 to 9 parts by weight 3 to 15 parts by weight 3 to 9 parts by weight 3 to 18 parts by weight 3 to 9 parts by weight 3 to 10 parts by weight wrinkle improving and moisturizing Composition for external application for skin.

6. The method of claim 5,
Wherein said composition inhibits hyaluronidase activity. &Lt; RTI ID = 0.0 > 11. < / RTI >