KR20140145709A - Extract of fermentative Zizyphus jujuba Fructus by Laetiporus sulphureus having lifespan extention of C. elegans via stress response and increasing collegen synthesis, and its cosmetic and therapeutic agent for wrinkle containing the same - Google Patents

Abstract

The present invention relates to a jujube-Laetiporus sulphureus fermented liquid which has effects of increasing longevity and resistance against heat stress and oxidative stress in C.elegans, and having collagen biosynthesis ability and to a drug and cosmetic material containing the same as an active ingredient for anti-aging and anti-wrinkling effects. The jujube-Laetiporus sulphureus fermented liquid of the present invention inhibits aging in C.elegans, thereby increasing the longevity. The jujube-Laetiporus sulphureus fermented liquid also has resistance against heat stress and oxidative stress, thereby leading to an outstanding antiaging activity and increases collagen synthesis, thereby suppressing wrinkling due to aging and being used for anti-wrinkling.

Description

{Extract of fermentative Zizyphus jujuba Fructus by Laetiporus sulphureus having lifespan extensions of C. elegans via stress response and increasing collegen synthesis, and its cosmetic and therapeutic agent for wrinkle containing the same}

The present invention relates to a cosmetic composition comprising a fermented jujube extract as an active ingredient, and more particularly to a cosmetic composition for anti-aging and wrinkle improvement by inoculating and fermenting a mycelia of red mildew.

The extent to which aging of cells affects the aging of individuals (humans, animals) is not yet known. It is well known that the telomere at the end of the chromosome is shortened each time it breaks down and the cell dies when it reaches its limit. Recently, there has been research that collagen, which is the main component of cell epilepsy, is reduced to 65% of the 20s when it is in their 80s and is involved in aging. There are also studies that abnormal protein accumulation due to intracellular ER stress is involved in aging.

In skin aging, the three major components of connective tissue, collagen and proteoglycan, are catabolized by intermolecular cross-linking or branching, resulting in changes in cell-extracellular matrix interactions. In addition, the decrease of the polar cells and the turnover of the epidermal cells (turn over, usually 28 days) are reduced from 70 to 40% of their 30s, causing skin dullness. And collagen, ceramide reduction is the main cause of skin wrinkles due to aging. It has been reported that the administration of estrogen improves the moisturization by increasing the synthesis of hyaluronic acid as well as the collagen increase of the skin (EL-Domyati M et al .: Exp Dermatol 11, 2002)

Oxidative stress is known to be an important cause of aging. Cellular stressors include reactive oxygen species called reactive oxygen species (ROS), such as peroxides, hydroxy radicals, and hydrogen peroxide. RNS (reactive nitrogen species) such as nitric oxide and peroxynitrite are known, and lipid peroxides are also known as oxidative stress inducers. These oxides are known to bind to guanine, a DNA base, to induce mutagenesis, leading to skin aging and cancer. It has been reported that it participates in the arachidonic acid cascade and causes vasoconstriction and platelet aggregation to develop as a risk factor for arteriosclerosis or Alzheimer's disease. Vitamin C and B12 are known to be water soluble and lycopene and beta-carotene are known to be lipid soluble.

However, there have been few reports on aging regulators using natural materials. Therefore, it is considered that the development of natural substance new drugs by deriving daf-16-activating natural molecules, which are the downstream of calorie restriction, will contribute greatly to improvement of quality of life such as inhibition of aging, improvement of metabolic diseases, immune function and stress resistance .

Korean Patent Publication No. 10-1027111 (Registration Date: Mar. 23, 2011) discloses cosmetics having excellent skin irritation mitigation and anti-inflammatory effects. The cosmetic contains fermented jujube leaf extract obtained by fermenting jujube leaf extract as an active ingredient Wherein the cosmetic is a cosmetic.

The present inventors intend to provide a natural extract which is applicable to skin cosmetics and which is harmless to human body and is highly safe, and a cosmetic composition containing it as an active ingredient.

It is another object of the present invention to provide a natural extract having excellent wrinkle-reducing effect and moisturizing effect, and a cosmetic composition containing the extract as an active ingredient.

Also, the jujube extract of the present invention is fermented with red mushroom to be resistant to heat stress and oxidative stress, and may be anti-aging.

The present invention provides a cosmetic composition characterized by comprising a mixture composed of a fermented jujube fermented with an extract of jujube having an anti-aging and wrinkle-improving effect as an active ingredient.

On the other hand, in the cosmetic composition of the present invention, the cosmetic composition may be anti-aging.

On the other hand, in the cosmetic composition of the present invention, the cosmetic composition may be for suppressing heat aging.

On the other hand, in the cosmetic composition of the present invention, the cosmetic composition may be for improving skin wrinkles.

On the other hand, in the cosmetic composition of the present invention, the cosmetic composition may be for skin moisturizing.

On the other hand, in the cosmetic composition of the present invention, the extract may be any one selected from the group consisting of water, an anhydrous or hydrated lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate and 1,3-butylene glycol And may be extracted using an extraction solvent.

Meanwhile, in the cosmetic composition of the present invention, the mixture may be added in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition.

Meanwhile, in the cosmetic composition of the present invention, the cosmetic composition may be in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, And spray. ≪ / RTI >

INDUSTRIAL APPLICABILITY The present invention can provide a natural extract which is applicable to skin cosmetics and which is harmless to the human body and has excellent safety, and a cosmetic composition containing it as an active ingredient.

Further, the present invention can provide a natural extract having excellent anti-aging, anti-aging effect, wrinkle-reducing effect and moisturizing effect, and a cosmetic composition containing it as an active ingredient.

Fig. 1 shows the collagen production effect of the jujube fermentation broth.
FIG. 2 shows the life extension effect of the jujube fermentation broth.
Fig. 3 shows the effect of heat stress resistance of the jujube fermentation broth.
Fig. 4 shows the oxidative stress resistance effect of the fermented broccoli fermentation broth.

The inventors of the present invention have conducted studies to prepare extracts from various natural materials as a raw material by various extraction methods and to select substances having excellent effects such as anti-aging effect and wrinkle-reducing effect. As a result, it was found that the extract prepared by culturing the jujube extract with the red mushroom was most suitable for the purpose of the present invention described above.

Jujube (Chinese date.) Is a botanical taxonomic tree belonging to the family Seagull and Jujube. It is a Chinese jujube (scientific name: Zizyphus jujuba Miller., English name: Chinese jujube) and Indian jujube (scientific name: Zizyphus mauritiana LA M., English name: Indian jujube). Dried in autumn, it is dried in the sun, its surface is reddish brown, elliptical, 1.5 to 2.5 cm long, and has a sweet taste when cooked red. The jujube leaves are homogenous, narrow ovate with dull mount and three large veins develop from the base. As a result, 2-3 flowers are bloomed and bloomed in June. Jujube has a beta-carotene ingredient that has the effect of reducing the active oxygen that activates aging. Jujube also helps the body drain unnecessary moisture and helps circulation.

To date, the jujube extract has been widely used for other purposes. However, as in the present invention, it has not been known whether the fermented form, that is, the fermented jujube fermented mushroom, has remarkably high anti-aging, wrinkle and moisturizing effect than when using the extract alone.

The cosmetic composition comprising the mixture of the jujube extract of the present invention obtained by fermenting with Laetiporus sulphureus CS0218 (KFCC 11494P) as an active ingredient has excellent anti-aging effect, .

In extracts of jujube, the active ingredient of the cosmetic composition of the present invention, the extracts can be prepared according to conventional methods known in the art, that is, under the conditions of ordinary temperature and pressure, using conventional solvents. That is, the jujuba extract, which is an effective ingredient of the composition of the present invention, is a solvent selected from the group consisting of water, an anhydrous or a lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate and 1,3- Or extracting jujube with the above-mentioned solvent, and then mixing them.

Among the above solvents, alcohols are preferable, and ethanol is more preferable. At this time, the ethanol is preferably 70% ethanol. The suitable amount of the extraction solvent is 1 to 15 times, more preferably 5 to 10 times, most preferably 7 times the jujuba dry weight.

On the other hand, the fermented broccoli fermentation broth of the present invention includes not only the extract obtained by the above extraction method, but also the extract obtained through ordinary purification process. For example, by separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatographies (made for separation by size, charge, hydrophobicity or affinity), and the like, It is also interpreted that the fraction is included in the fermented broccoli fermentation broth of the present invention.

In addition, the fermented jujube fermented jujube of the present invention may be produced in powder form by an additional process such as vacuum distillation, freeze-drying, or spray drying.

According to a preferred embodiment of the present invention, the content of the jujuba fermentation broth added to the cosmetic composition is 0.0001 to 30.0% by weight, preferably 0.001 to 15.0% by weight, and most preferably 0.1 To 10.0% by weight. When the content of the fermented broccoli fermented juice is less than 0.0001% by weight, the effect of improving the anti-aging and wrinkling is insignificant. When the content is more than 30%, the effect of increasing the content is not increased and it is not economical.

Meanwhile, the cosmetic composition of the present invention may contain, as an effective ingredient, the components commonly used in cosmetic compositions in addition to the fermented jujube fermented mushroom fermentation product, and may contain conventional components such as antioxidants, stabilizers, solubilizers, vitamins, An antioxidant, an anti-inflammatory agent, and a carrier.

Meanwhile, the cosmetic composition of the present invention may be prepared in any form conventionally produced in the art, and examples thereof include a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, Containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays. However, the present invention is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a convergent lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

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

When the formulation of the cosmetic composition of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, Propane / butane, or dimethyl ether. ≪ RTI ID = 0.0 >

When the formulation of the cosmetic composition of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate , Propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the cosmetic composition of the present invention is in the form of a suspension, the carrier component may include water, a liquid diluent such as ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester Suspending agent, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the cosmetic composition of the present invention is a cleansing composition containing an interfacial active agent, it is preferable to use, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, Fatty acid amide ether sulfate, alkylamido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

On the other hand, the fermented broccoli fermentation broth of the present invention exhibits an anti-aging effect (see Experimental Example 3 and Experimental Example 5), a heat aging inhibitory effect (Experimental Example 4) and a wrinkle-improving effect (Experimental Example 2) . In addition, the cosmetic composition prepared by containing the jujube red mandarin mushroom having the above effect as an active ingredient showed excellent skin moisturizing effect. (Reference Example 6)

Hereinafter, the present invention will be described in more detail with reference to the following Production Examples. However, the following Production Examples are intended to further illustrate the present invention, and the scope of the present invention is not limited to the following Production Examples.

[ Manufacturing example  One: Jujube red duck mushroom  Preparation of fermentation broth]

Jujube was washed with purified water, dried and then crushed and made fine using a sieve of 100 mesh. To this, 100 g / L of jujube juice was mixed with purified water and added to the culture medium of Laetiporus sulphureus CS0218 (KFCC 11494P). 1 ~ 4% of glucose was added as a carbon source, and 0.2 ~ 0.5% of peptone was further added thereto. The cultures were incubated for 7, 14 days at 37 ° C, pH 5-7 using a 5 L fermenter. After culturing, the culture was centrifuged to remove the cultures first.

After the fermentation, the fermented jujube was firstly removed from the fermented juice, and the ethanol was added to the final 70% (v / v) ethanol aqueous solution. The mixture was refluxed three times for 5 hours, cooled, and filtered through Whatman # 10 filter paper Respectively. The thus obtained filtrate was finally filtered using a 0.25 uM filter. When the filtration is completed, the extract obtained in the extraction step is transferred to a concentration tank and concentrated under reduced pressure or freeze-dried at 50 ° C or lower. The mixed extract was prepared by using at least one solvent selected from purified water, ethanol, butylene glycol and propylene glycol so that the reduced pressure concentrate or lyophilized product contained 0.001 to 70.0 wt%.

[compare Manufacturing example  1: Preparation of jujube extract]

The same jujitsu as in Preparation Example 1 was washed with purified water, dried, and then pulverized and finely ground using a sieve of 100 mesh. 70% ethanol was added to 100 g / L of jujube crushed product, and the mixture was refluxed three times for 5 hours. After cooling, it was filtered with a filter of whatman # 10. The thus obtained filtrate was finally filtered using a 0.25 uM filter. The filtered extract was concentrated under reduced pressure and freeze-dried at 50 DEG C or lower. A mixed extract was prepared by using at least one solvent selected from purified water, ethanol, butylene glycol, and propylene glycol so that the reduced pressure concentrate and the lyophilized product contained 0.001 to 70.0 wt%.

[ Experimental Example  One: Jujube red duck mushroom  Evaluation of cell viability of fermentation broth]

To evaluate the toxicity of the fermented jujube extract and jujube extract obtained in Preparation Example 1 and Comparative Preparation Examples 1 and 2 on cells.

Cytotoxicity was performed by modifying Mosmann's method of measuring cell viability using 3- (4,5-dimethylthiazol-2-yl) -2-5-diphenyltetrazolium bromide (MTT) reagent.

HDF was dispensed into 96-well plates at a concentration of 1 × 10 ⁵ cells / well and cultured at 37 ° C and 5% CO ₂ for 24 h. After the culture medium was removed, the sample was cultured for 24 h in a concentration-treated medium, and then the medium was removed and washed twice with PBS (phosphate buffered saline). MTT was dissolved in PBS at 300 μg / mL and 100 μL was added and incubated at 37 ° C and 5% CO ₂ for 4 h. 150 μL of DMSO was added to each well, stirred for 30 min, and absorbance was measured at 570 nm.

As a result, no cytotoxicity was observed at all concentrations of each sample. From this, it can be confirmed that the fermented jujube leaf extract of the present invention is harmless to the human body and has excellent safety.

[ Experimental Example  2: Jujube red duck mushroom  Evaluation of collagen production effect of fermentation broth]

Collagen aggregation performance was assessed by the use of antibodies that recognize the C-terminus of the collagen precursor (PIP, Procollagen Type 1 C-peptide). The jujube extract, the jujube extract fermentation 7 days, and the fermentation 14 day extract were treated with 100 ug / mL 200 ug / mL, respectively. The culture broth was collected and the amount of C-terminal end of the collagen precursor was measured.

According to the results shown in Fig. 1, the amount of collagen produced in the jujube extract (zjf-100, -200) was not different from that of the control group, and the change in the amount of 100 ug / mL of the jujube fermented on the 7th day of fermentation was not observed . On the 7th day of fermentation, statistically significant activity was observed at the concentrations of F7-100 (100 ug / mL) and F7-200 (200 ug / mL) and on the 14th day of fermentation (F14-200) at 200 ug / mL.

Therefore, it can be confirmed that the culture of jujube red mandarin mushroom of the present invention has an excellent effect for improving wrinkles.

[ Experimental Example  3: Jujube red duck mushroom  Evaluation of life extension effect of fermentation broth]

Experiments were carried out using wild type N2 C. elegans (Caenorhabditis Genetics Center, University of Minnesota, Minneapolis, MN, USA). Experiments were carried out at 20 ℃ and 25 ℃ in groups of drug - treated and untreated groups. The adult worms were transferred to a new plate and counted dead worms.

2, the jujube extract (zjf-200 ug / mL) showed no life extension effect at 20 ° C and 25 ° C. In the group treated with 7-day fermentation of jujube extract (F7-zjf-200 ug / mL), there was no life-extension effect in the group cultured at 20 ° C. However, in the stress-induced condition at 25 ° C., Which was significantly prolonged. In the group treated with jujube fermentation extract (F14-zjf-200ug / mL), no effect of life extension was observed in the group cultured at 20 ° C. However, in the case of inducing stress at 25 ° C., Which was significantly prolonged.

Therefore, it was confirmed that the culture of jujube red duck fungus of the present invention has a remarkable effect on life prolongation using small nematodes.

[ Experimental Example  4: Jujube red duck mushroom  Evaluation of heat stress resistance effect of fermentation broth]

One hundred nematodes of the Syncronized L1 stage were transferred to an NGM agar plate and cultured for 72 hours to become L4 stage Lavae. Ten adult worms were transferred to a 6 cm plate treated with the drug and an untreated plate, respectively, and the adult worm was removed from the plate for 3 hours. During the reproductive period, the drug was transferred to a fresh medium that had been treated daily, and on the sixth day, thermal stress was given for 8 hours in a 35 ° C incubator. The number of dead nematodes was counted and compared with the control group.

According to the results shown in Fig. 3, the treatment group of 200 ug / mL of jujube fermented with fermented jujube extract showed a 12% survival rate higher than that of the control group, indicating that the heat stress resistance was significantly (t = 4.017). The Jonckheere-Terpstra test was conducted to determine if there was any change in the efficacy of the drug during the fermentation period, but the efficacy of the drug did not increase or decrease with fermentation period.

Therefore, it was confirmed that the culture of jujube red mandarin mushroom of the present invention has an excellent effect on the heat stress resistance.

[ Experimental Example  5: Jujube red duck mushroom  Fermentation Oxidative  Evaluation of stress resistance effect]

C. elegans According to aging, H₂O₂ It is known that the associated ROS levels tend to increase. To investigate the relationship between oxidative stress resistance and inhibition of aging,₂O₂Were carried out. One hundred nematodes of the Syncronized L1 stage were transferred to an NGM agar plate and cultured for 65 hours to become L4 stage Lavae. Syncronized L4 stage nematodes were transferred to a 6-cm plate and a drug-untreated plate, respectively, in an amount of 10-15 eggs. The eggs were laid for 3 hours and the adult worm was removed from the plate. During the reproductive period, the cells were transferred to a fresh medium treated with the drug each day and transferred to M9 buffer containing 0.8 mM hydrogen peroxide on the 6th day for 8 hours to measure oxidative stress.

According to the results shown in Fig. 4, the 200 ug / mL of the fermented 7-day liquid and the 100 ug / mL of the fermented 14-day liquid were significantly higher in the oxidative stress resistance than the control. The Jonckheere-Terpstra test was performed to determine if the effect of the drug on the fermentation time was changed. As a result of the analysis, it was statistically confirmed that the magnitude of effect did not change according to the effective date.

Therefore, it was confirmed that the culture of jujube red duck fungus of the present invention has an excellent effect on the oxidative stress resistance.

[ Manufacturing example  2: Jujube red duck mushroom  Fermented Cosmetic  Produce]

A cosmetic composition containing 10.0% by weight of the fermented jujube fermented in a fermented product of Preparation Example 1 was prepared according to the composition shown in the following Table 4, referred to as Prescription Example 1, and a cosmetic composition containing no Nevaneve complex was prepared, Example 2.

ingredient Prescription Example 1
Content (% by weight) Comparative Prescription Example 1
Content (% by weight) Production Example 1 10.0 - Comparative Preparation Example 1 - 10.0 1,3-BG 10.0 10.0 glycerin 5.1 5.1 Propylene glycol 4.2 4.2 Tocopheryl acetate 3.0 3.0 Liquid paraffin 4.6 4.6 Triethanolamine 1.0 1.0 Squalane 3.1 3.1 Macadamia nut oil 2.5 2.5 Polysorbate 60 1.6 1.6 Sorbitan sesquiterate 1.6 1.6 Propylparaben 0.6 0.6 Carboxyl vinyl polymer 1.5 1.5 incense a very small amount a very small amount antiseptic a very small amount a very small amount Purified water Balance Balance system 100 100

[ Experimental Example  6: Jujube red duck mushroom  The skin of the fermentation liquid Moisture power  Improvement effect]

In order to examine the effect of the cosmetic composition containing the fermented jujube fermented jujube extract of the present invention on the skin moisturizing ability, the following experiment was conducted. Twenty patients in the 20s to 40s without skin diseases were divided into two groups each consisting of 20 patients per group. The nutritional cream of the cream of the prescription example 1 and the comparative prescription examples 1 to 2 prepared in the above Preparation Example 1 Twice a day for one month. The skin conductivity was measured by using a corneometer (CM820 courage Khazaka electronic GmbH, Germany) under the constant temperature and humidity conditions (24 ° C, 40% humidity) before the start of application, and the skin conductivity was measured at 1 week, 2 weeks, The rate of increase in skin conductivity (%) was measured and the rate of increase was evaluated. The results are shown in Table 1 below.

sample After 1 week after 2 weeks After 4 weeks Prescription Example 1 55 61 66 Comparative Prescription Example 1 23 31 38

As shown in the results of Table 1, in the case of Formulation Example 1, which is a cosmetic containing 7-day fermentation broth of Daechu red mildew of the present invention, the rate of increase in skin conductivity was much higher than Comparative Prescription Examples 1 and 2. Since the skin conductivity is generally proportional to the skin moisture content, the above results confirm that cosmetics containing jujube red mandarin mushrooms maintain their skin moisture content higher than those without cosmetics. In addition, there was no difference in the moisture content of the skin when the fermentation period was set to 7 days and that when the fermentation period was set to 14 days, confirming that there was no difference in the effect according to the fermentation period.

Claims (8)

A cosmetic composition characterized by comprising an extract of a jujube having an anti-aging effect and a wrinkle-improving effect, which is fermented with red mushroom, as an active ingredient.
The method according to claim 1,
In the cosmetic composition,
The cosmetic composition is characterized in that it is for preventing skin aging.
The method according to claim 1,
In the cosmetic composition,
Wherein the cosmetic composition is for inhibiting thermal aging.
The method according to claim 1,
In the cosmetic composition,
Wherein the cosmetic composition is for improving skin wrinkles.
The method according to claim 1,
In the cosmetic composition,
Wherein the composition is for moisturizing the skin.
3. The method according to claim 1 or 2,
The above-
A cosmetic composition characterized by being extracted using any one extraction solvent selected from the group consisting of water, anhydrous or lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate and 1,3-butylene glycol .
The method according to claim 1,
The mixture may contain,
Is added in an amount of 0.0001 to 100.0% by weight based on the total weight of the cosmetic composition.
The method according to claim 1,
In the cosmetic composition,
The composition has any one of the formulations selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansers, oils, powder foundations, emulsion foundations, wax foundations and sprays ≪ / RTI >

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