KR101904971B1 - A solid fermentation composition from trichosanthis and vegetable worms and a functional cosmetic composition comprising the same - Google Patents

Abstract

The present invention relates to a solid fermentation product of Trichosanthis spp. And a vegetable worm, and a functional cosmetic composition containing the same. More specifically, the present invention relates to a solid fermentation product obtained by inoculating Cordyceps sinensis A solid fermentation product obtained by solid fermentation, and a functional cosmetic composition for promoting antioxidation, skin aging prevention and skin regeneration comprising the same. The solid fermented product of the present invention is a composition derived from a natural product having no cytotoxicity and is excellent in antioxidation, inhibition of skin aging and promotion of skin regeneration, and is useful as an antioxidant and anti-aging agent, particularly a functional cosmetic for skin regeneration and wrinkle improvement It is very useful in the cosmetics industry.

Description

FIELD OF THE INVENTION The present invention relates to a solid fermented product of Cordyceps militaris and a functional cosmetic composition comprising the same, and a functional cosmetic composition comprising the same. BACKGROUND ART < RTI ID = 0.0 >

The present invention relates to a solid fermentation product of Trichosanthis spp. And a vegetable worm, and a functional cosmetic composition containing the same. More specifically, the present invention relates to a solid fermentation product obtained by inoculating Cordyceps sinensis A solid fermentation product obtained by solid fermentation, and a functional cosmetic composition for promoting antioxidation, skin aging prevention and skin regeneration comprising the same.

Skin is a part of the body that is directly exposed to the external environment. It is exposed to ultraviolet rays caused by exposure to sunlight in daily life, air pollutants such as yellow dust or automobile soot, and various bacteria, thereby causing skin trouble and irritation. . Such aging of the skin reduces the elasticity of the skin on the appearance of the skin to form wrinkles or to form spots and senile spots to cause skin pigmentation.

In general, aging of the skin is accompanied by various theories such as somatic mutation, excessive production of free radicals, autoimmune reaction, accumulation of toxic substances, errors generated during gene replication, transformation of dendritic proteins, And the diseases caused by various diseases such as adult diseases are attributed to reactive oxygen species (ROS) is recognized that the active oxygen species originating from oxygen and nitrogen to control the natural antioxidants to develop active research is progressing have. The reactive oxygen species, usually known as harmful oxygen, is a lipid peroxide such as peroxide anion radical, hydrogen peroxide, hydroxy radical, peroxidized lipid or free radical generated in the most stable form, It occurs even under normal conditions, but the presence of antioxidant enzymes such as superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx) and the like inhibit the production of excessive reactive oxygen species. However, when the active oxygen species is abnormally excessive due to stress and external environmental factors and is not eliminated, the oxidative stress due to the free radicals is added to the living body and DNA, RNA, protein, Cell membrane and cell structure damage, cancer, tissue damage, and aging.

Studies on antioxidants capable of controlling or eliminating these active oxygen species have been actively conducted in order to suppress the rapid increase of various diseases and aging in the modern age. Studies on the efficacy of natural antioxidants such as tocopherol, ascorbic acid, carotenoids, flavonoids and glutathione, and their stabilization techniques, and studies on the synthetic antioxidants butylated hydroxytoluene (BHT), butylated hydroxyanisol (BHA), and many other antioxidants have been continuously studied. Among these antioxidants, tocopherol has a relatively low antioxidative effect, and synthetic antioxidants such as BHA and BHT, which are highly effective, are used in pharmaceuticals and foods, but there is a point of mutagenicity and toxicity, and in the case of ascorbic acid, Development of the obtained derivatives and formulations is progressing, and development of a natural antioxidant material having more stable and effective effect is urgently required.

In order to solve such a problem, many studies have been made on a cosmetic composition using an extract which is relatively low in toxicity, safe and effective for skin, among phytochemicals, It has been required to develop a material which maximizes its efficacy since the effect of the ingredient is limited.

On the other hand, in the case of natural extracts through fermentation, sugar components present in natural products are converted into alcohols as carbon sources in the metabolic process of microorganisms without using harmful organic solvents, so that the active ingredients in natural products, especially flavonoids, Polyphenols, catechins, etc. are present as glycated and stable compounds, and the extractability of the active ingredient is increased through fermentation, and excellent skin efficacy is expected by the active ingredient which is excluded from sugar and activated.

Cordyceps is a mushroom in the winter and a mushroom in the summer. The spores and mycelium of the fungus, a kind of fungus, invade into living insects and lurk during winter. Is an odd mushroom that causes insects to die in summer and uses endogenous materials as a nutrient source to produce endosynthetic sclerotia and then to form fruit bodies outside the body of insects.

There have been more than 750 kinds of Cordyceps in the world so far and more than 100 species such as Cordyceps spp., Paecilomyces spp, Isaria spp. do.

According to Chinese ancient books, Cordyceps is not only effective in cancer, liver disease, lung disease, brain disease, cardiovascular disease, heart disease, diabetes, etc., but it also has pulmonary tuberculosis, elderly debilitating chronic tuberculosis anemia, . Recently, it has been reported that tumor suppression rate is 83%, which is an anticancer effect and antidote to drug poisoning, and it has natural healing power and it is reported that recovery is effective when heavy physical exercise is heavy.

It is also used as a name for the roots of the sky, as it is dried in the sun with the peeled roots of Trichosanthis kirillowii Max. Or T. killillowii Max. Var Japonicum KITAMURA.

It is a perennial herbaceous herb of surgery. It is a perennial herbaceous plant root of perennial herbaceous plants (baconaceae). It grows mainly in the southern part of the country, especially in Jeju Island, northern mountainous regions or under forests. It is an uneven pale yellowish-white cylinder with a diameter of 3 ~ 5㎝ and a length of 5 ~ 10㎝. It is often split vertically and has irregular yellowish brown vascularity. There is no smell and the taste is a bit red. The cut surface is light yellow and slightly fibrous. The cross section of the magnifying glass shows wide radiating texture and yellow spots or small holes (conduits).

It is used in children's skin diseases such as fever, broad-spectrum, disinfectant, jaundice, diabetes, stroke or spray. It is also used as a contraceptive in China.

KR 10-2013-0149221 A KR 10-2007-0038920 A KR 10-2007-0091382 A

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made in order to solve the problems of the prior art as described above, and an object of the present invention is to provide a method for producing a solid Fermented solid fermented product and a functional cosmetic composition using the same.

Disclosure of the Invention The present invention has been conceived in order to solve the above-mentioned problems, and provides a solid fermented product of Pseudomonas aeruginosa and Cordyceps sinensis produced by inoculating and cultivating a mycelia of Cordyceps mushroom on sterilized Pseudomonas aeruginosa.

The fungus is coordinating sepseu sinen system (Cordyceps sinensis), Cody sepseu Milla other leases (Cordyceps militaris), Cody sepseu Scarab Cola (Cordyceps scarabaecola), Cody sepseu press Tansu (Cordyceps nutans), Cody sepseu seupeko Seppala (Cordyceps sphecocephala) , Cordyceps tricentri , Paecilomyceps tenuipes , and Paecilomyces japonica . ≪ IMAGE >

The culture is preferably performed under conditions of a temperature of 20 to 35 DEG C, a relative humidity of 70 to 90%, and a culture period of 7 to 45 days.

In addition, the present invention provides a cosmetic composition comprising the solid fermented product of the present invention and the cucumber herb.

The cosmetic composition preferably has a function selected from the group consisting of antioxidation, skin aging prevention, and skin regeneration promotion.

The cosmetic composition of the present invention can be used in cosmetics such as a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a powder foundation, an emulsion foundation, a wax foundation, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, , A lipstick, a shadow, a shampoo, and a rinse.

The solid fermented product of the present invention is a composition derived from a natural product having no cytotoxicity and is excellent in antioxidation, inhibition of skin aging and promotion of skin regeneration, and is useful as an antioxidant and anti-aging agent, particularly a functional cosmetic for skin regeneration and wrinkle improvement It is very useful in the cosmetics industry.

Fig. 1 shows a process for producing a solid fermented product of the present invention.
Fig. 2 shows the SOD radical scavenging activity of the solid fermented product of the present invention.
Fig. 3 shows the Xanthine oxidase assay results of the solid fermented product of the present invention.
FIG. 4 is a northern blot diagram showing the expression pattern of SIRT1 (Sirtuin1) gene in the fermented product of P. vannamei of the present invention.
FIG. 5 is a graph showing the expression pattern of SIRT1 (Sirtuin1) gene in the fermented product of Chinese cabbage caterpillar fungus of the present invention.
Fig. 6 shows the skin wound regeneration effect of the solid fermented product of the present invention on the time course.
Figure 7 shows the NRU phototoxicity results of CPZ (chloropromazine), known as a phototoxic substance.
Figure 8 shows the NRU phototoxicity results of the solid fermented caterpillars of the present invention.
Fig. 9 shows the MTT assay results of the solid fermented product of the present invention.

Hereinafter, the present invention will be described in detail.

The inventors of the present invention have conducted extensive research to develop materials for functional cosmetic compositions using natural products. As a result, the present inventors found that the solid fermented product of the ginseng extract obtained from the step of inoculating the mycelium of the mushroom with the mushroom, Skin aging and skin regeneration activity, and suggest that the solid fermented product of the present invention can be used as a functional cosmetic composition.

Accordingly, the present invention provides a solid fermented product of the ginseng root and Cordyceps, which is prepared by inoculating and culturing the mycelia of Cordyceps melliferae with sterilized gull roots.

The Cordyceps sinensis according to the present invention is a fungus belonging to the Cordyceps mellifera of the Acanthopanax senticosus, and specifically includes Cordyceps spp., Paecilomyces spp., Isaria spp. ( Phytocordyceps spp.), And more than 750 species are known worldwide.

In the present invention, all of these Cordyceps sinensis can be used, and preferably Cordyceps sinensis , Cordyceps militaris , Cordyceps scarabaecola , Codisps mutans, Cordyceps nutus , Cordyceps sphecocephala , Cordyceps tricentri , Paecilomyceps tenuipes , and Paecilomyces japonica . , More preferably Paecilomyces japonica, also referred to as Snowcone Cordyceps.

The mycelium of Cordyceps mellifera of the present invention can be produced by seed culture of Cordyceps spp.

As a preferable example, the mycelia of Cordyceps sinensis are inoculated into a PDA (Potato Dextrose Agar) medium, and then subcultured at about 4 weeks intervals at a temperature of about 25 ° C. .

The seed culture is a process for culturing the stored mycelium of Cordycepian mycelium prior to the main culture. In a preferred embodiment, the seed culture can be obtained as a liquid culture by inoculating seed culture, i.e., mycelium, into a culture medium for mushroom mycelium, and then culturing the culture in a rotary shake.

The roots of the present invention refer to roots that have been peeled off. For example, Trichosanthis kirilowii or T. radix may be used as the plant of the genus Trichosanthis spp. Lt; / RTI >

The sterilization of the ganglioside can be achieved by washing the prepared ganglioside, dehydrating and drying it, and applying various known sterilization methods. In a preferred embodiment, the sterilization can be autoclaved at 121 DEG C for 30 minutes.

The method for inoculating the mycelia of the present invention with the sterilized baculovirus is aseptically inoculated into a sterilized baculovirus by a predetermined amount of the liquid culture obtained by the baculovirus culture described above.

The cultivation is preferably performed at a temperature of 20 to 35 ° C, a relative humidity of 70 to 90%, and a cultivation period of 7 to 45 days. As a preferred example, the cultivation is carried out at a temperature of 25 ° C In an incubator with a relative humidity of 80% for 30 days.

The fermented solid fermented celery root celeriac is a form in which the surface of the celery root is entirely covered with the mycelia of the celery root, thereby making it possible to produce an extract of the solid fermented celery root celeriac.

The solvent used for the extraction may be water or lower ethanol, but is preferably hot water extraction. As a preferred embodiment, the supernatant can be taken by adding about 5 to 150 times the volume of hot water to the obtained fermented ginseng root of Cordyceps sinensis, shaking for 1 to 5 hours, and centrifuging. The above process may be performed 1 to 5 times, preferably 3 times.

The solid fermented product of the present invention or extract thereof can be prepared by the above-mentioned concrete manufacturing process, and it can be applied as the material of the functional cosmetic composition described below.

Accordingly, the present invention provides a functional cosmetic composition containing the fermented product of the present invention.

The term " functional " means an activity selected from the group consisting of antioxidation, prevention of skin aging, and promotion of skin regeneration.

The solid fermented product of Cordyceps militaris as a main component of the cosmetic composition of the present invention may be contained in an amount of 0.01 to 50% by weight, preferably 0.1 to 20% by weight, more preferably 1 to 10% by weight based on the total weight of the composition . It is possible to secure appropriate formulation stability in such a content range and to anticipate the effect of the desired antioxidation, prevention of skin aging and promotion of skin regeneration.

The composition of the present invention may further include known natural extracts or other ingredients for the antioxidation, prevention of skin aging and promotion of skin regeneration, in addition to the solid fermented product of the Chinese cabbage root, .

In addition, the composition may also contain components commonly added to cosmetic compositions, such as fatty substances, organic solvents, solubilizers, thickeners, gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, A cosmetic or dermatological composition such as a perfume, a perfume, a surfactant, water, an ionic or nonionic type emulsifier, a filler, a sequestering and chelating agent, a preservative, a vitamin, a barrier, a wetting agent, an essential oil, a dye, a pigment, a hydrophilic or lipophilic active agent May be formulated into a particular formulation, including adjuvants or carriers commonly used in the field of dermatology.

The cosmetic composition of the present invention may be prepared in any form conventionally produced in the art and may be formulated into various forms such as solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, Containing cleansing, oil and spray, but are not limited thereto. More specifically, the present invention relates to a cosmetic composition for cosmetics such as a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a powder foundation, an emulsion foundation, a wax foundation, a cleansing cream, a cleansing foam, a cleansing water, Rose, lipstick, shadow, shampoo, and rinse.

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

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as the carrier component. In particular, in the case of a spray, a mixture of chlorofluorohydrocarbons, 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, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments. The following examples illustrate one preferred embodiment of the present invention and are not to be construed as limiting the scope of the present invention.

[Example]

1. Preparation of sample

1.1. Preparation of Hot Water Extract of Fermented Solid Fermented Cordyceps

Paecilomyces Japonica was cultivated using a PDA (Potato Dextrose Agar, Difco, Co., USA) medium at 25 ° C using a strains under cultivation in Palm Bios Co., . In a 500 ml flask, 100 ml of mushroom mycelial solid medium (1 cm x 1 cm, 3 pieces) was inoculated with the mycelia of the snowy Chinese mushroom mycelium in a volume of 100 ml, and then inoculated in a rotary shaking incubator (Daihan Labtech Co.Korea, 25 ± 1 ° C, 130 rpm) Day, and used as liquid seeds.

In the 3L culture vessel, 500 g of ullulose was sterilized at 121 캜 for 30 minutes, and then cooled at room temperature. Then, 60 ml of liquid seedlings were aseptically inoculated into 500 g of sterilized gull root. Fermentation was carried out for 30 days in an incubator at 25 ° C and 80% relative humidity. After fermentation, 10 times volume of hot water was added, and the mixture was shaken for 2 hours and then immersed for 72 hours. After centrifugation at 10,000 rpm for 10 minutes, the supernatant was taken. After repeating this three times, the extract was concentrated under reduced pressure to prepare a powder Respectively.

1.2. Preparation of Hot Water Extract of Solid Fermented Mushrooms

The hot-water extract of Lactobacillus japonica solid fermented product was prepared in the same manner as in item 1.1 except that the mushroom was used in place of the moth clown-grass seedlings.

1.3. Preparation of Hot Water Extract of Fermented Juice of Ganoderma Lucidum

The hot water extract of the fermented mushroom ganoderma lucidum was prepared in the same manner as in item 1.1 above except that the mung bean was used instead of the ginseng cucumbers.

2. Activity evaluation

2.1. Antioxidant activity verification

The antioxidant activity of the fermented product of the above-prepared ginseng celeriac / situ mushroom / gingko mushroom was evaluated. (1, 1-diphenyl-2-picryl hydrazyl) radical scavenging activity and FRAP activity were measured in the solid fermented product (0.5 mg / ml)

For DPPH scavenging ability, 380 μl of 2 × 10 ^-4 M DPPH solution dissolved in 99.5% ethanol was added to 20 μl of diluted samples at various concentrations, and the mixture was reacted at 37 ° C. for 30 minutes. Then, microplate reader (Asys Hitech, Expert 96, Asys Co., Austria). DPPH free radical scavenging activity was expressed as a percentage as shown in Table 1 below.

[Table 1] DPPH radical scavenging activity

The FRAP activity was carried out according to the known method for the prepared fermented caterpillar fungus / situ mushroom / Ganoderma lucidum solid fermentation (0.5 mg / ml).

[Table 2] FRAP activity

2.2. Cell Regeneration / Aging Suppression Test

2.2.1. Elastase inhibitiory assay

The results of evaluating the elastase inhibitory activity (%) for the prepared fermented celecoxib / mushroom / gingko mushroom are shown in Table 3 below.

[Table 3] Elastase inhibitory assay

Based on the above results, elastase inhibitory activity (%) was measured by concentration (30, 100, 300 and 1,000 μg / ml) and compared with positive control EGCG (epigallocatechin gallate) I could.

2.2.2. Superoxide dismutase assay

The SOD inhibitory activity of three solid fermented products tested by DPPH and FRAP and Elastase inhibitiory assays showed that SOD inhibition activity was higher than that of the extracts fermented with Chinese cabbage and mushroom at 30 ㎍ / (Fig. 2).

2.2.3. Xanthine oxidase assay

As a result of xanthine oxidase assay of three solid fermented products, xanthine oxidase activity tended to decrease according to the concentration of the extract of Wuchuan root extract (Fig. 3). In addition, the activity of xanthine oxidase, similar to that of raw materials, was also decreased in the fermentation broth of. These results suggest that the xanthine oxidase inhibitory activity by the fermentation of Rumen root is generally decreased in a concentration - dependent manner.

2.2.4. Identification of SIRT1 gene expression

SIRT1 has been shown to increase the lifespan by delaying proliferative aging by altering transcriptional activity to restore DNA damage, with resistance to oxidative attack while inhibiting apoptosis and proliferation through deacetylation. By confirming the expression level of SIRT1 gene, we confirmed that the fermented product of. 근 is effective for aging. Positive control used resveratrol, which is known to activate the SIRT1 gene.

As shown in FIGS. 4 and 5, when the degree of SIRT1 gene expression was measured in a fermented product of Cordyceps sinensis, it was lower than that of resveratrol but higher than that of untreated group. In particular, the expression level of SIRT1 gene Respectively. From the above results, it is expected that the solid fermentation of Bacillus subtilis induces the change of the natural material component, thereby increasing the expression of the SIRT1 gene and thus the anti-aging effect.

2.2.5. Wound healing assay

The wound healing assay was performed on human fibroblast BJ cells. BJ cells were not completely healed after 14 hours of treatment with the extracts of Bombyx mori, but showed some therapeutic effect. On the other hand, the solid fermented product of Cordyceps sinensis showed increased degree of wound healing compared with the extract of Walnut root. These results show that the results of the elastase inhibitory activity showed a tendency similar to that of the solid fermented product of P. vannamei, showing excellent elastase inhibitory activity and excellent effect on STIR1 gene expression, and these results are related to each other (FIG. 6).

2.3. (3T3 NRU phototoxicity test)

The NRU phototoxicity of the bovine leucocyte solid fermentation was measured using CPZ (chloropromazine) known as a phototoxic substance as a positive control using NIH / 3T3 cells (FIGS. 7 and 8). The MPE value and the PIF value of the extracts of Culexopsis lanceolata were -0.069 and 1.000, respectively. The MPE values were within the range of MPE <0.1 and PIF <2, It was judged to be absent. However, the MPE value of mushroom was 0.141 and the PIF value was 1.000. The MPE value of the mushroom was 0.127, the PIF value was 1.000 and the MPE value was in the range of 0.1 <MPE <0.15. . The above results confirm that even when the solid fermentation is performed using the Cordyceps, the phototoxicity is not observed when the Cordyceps is used.

2.4. Cytotoxicity test (MTT assay)

In order to evaluate the cytotoxicity of the fermented product of Bacillus subtilis, cell viability (%) was measured after MTT assay using BJ cells, and the results are shown in FIG. Cell viability decreased to less than 80% at a concentration higher than 300 μg / ml. In the case of the solid fermented product, the cell viability of the mushroom fermented product tended to decrease as the concentration increased. However, the cell viability of the mushroom fermented product and the mushroom fermented product was found to be 80% or more even at the treatment concentration of 300 μg / ml.

Claims (6)

A functional cosmetic composition for promoting skin regeneration, comprising a solid fermented product of ginseng root and Cordycepsus prepared by inoculating and culturing the mycelium of Cordyceps mellifera to sterilized gulloin roots. The method of claim 1, wherein the fungus is coordinated sepseu sinen sheath (Cordyceps sinensis), coordination sepseu ms other less (Cordyceps militaris), coordination sepseu Scarab coke (Cordyceps scarabaecola), coordination sepseu press Tansu (Cordyceps nutans), coordination sepseu seupeko Seppala (Cordyceps sphecocephala), coordination sepseu tree Sentry (Cordyceps tricentri), L indeed My process Te sister Fes (Paecilomyceps tenuipes) and L indeed functional cosmetic composition being selected from the group consisting of my process japonica (Paecilomyces japonica) . The functional cosmetic composition according to claim 1, wherein the culture is performed at a temperature of 20 to 35 DEG C, a relative humidity of 70 to 90%, and a culture period of 7 to 45 days. The cosmetic composition according to claim 1, wherein the functional cosmetic composition is at least one selected from the group consisting of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a powder foundation, an emulsion foundation, a wax foundation, a cleansing cream, a cleansing foam, a cleansing water, , Powder, fact, lip gloss, lipstick, shadow, shampoo and rinse. delete delete

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