KR20190046246A - Fermented composition of ice plant and collagen for anti-aging and anti-wrinkle, method for preparing the same and cosmetic composition comprising the same - Google Patents

Abstract

The present invention relates to a mixed fermented product of collagen, a collagen, a method for producing fermented seafood, and a cosmetic composition comprising the same, and more particularly, to a cosmetic composition comprising a non-fermented raw material To a complex fermented product of an ice plant and collagen having increased skin aging and skin wrinkle improving effect, a process for producing the same, and a cosmetic composition containing the same.
The ice plant and collagen-based fermented product and fermented seafood according to the present invention exhibit remarkably improved skin aging-preventing effect and skin wrinkle-improving effect as compared with non-fermented product, and thus can be very usefully used for manufacturing functional cosmetics.

Description

TECHNICAL FIELD The present invention relates to a fermented seaweed, a fermented seaweed, and a cosmetic composition comprising the same, and a cosmetic composition comprising the same, method for preparing the same and a cosmetic composition comprising the same.

The present invention relates to a mixed fermented product of collagen, a collagen, a method for producing fermented seafood, and a cosmetic composition comprising the same, and more particularly, to a cosmetic composition comprising a non-fermented raw material To a complex fermented product of an ice plant and collagen having increased skin aging and skin wrinkle improving effect, a process for producing the same, and a cosmetic composition containing the same.

Fermentation refers to the process by which microorganisms possess enzymes that change raw materials and produce beneficial components. During the fermentation process, active ingredients are extracted and the content of functional materials increases.

And the fermentation technology has the meaning of biologic conversion, biosynthesis, biocatalyst, etc., Meaning semantically redundant, and refers to the technology to produce desired products from precursors by using the enzymatic functions of microorganisms. Therefore, The fermentation technology used in the cell or enzyme stage is a major process constituting the present biotechnology industry with the main purpose of producing the useful substance. If one-dimensional fermentation technology that utilizes microorganisms that naturally occur in fermented foods is used, fermentation cosmetics require advanced technology, and if conventional fermentation methods are used for first generation fermentation, then second generation fermentation techniques for practical application of efficacious substances The bio-conversion fermentation technology that laid the foundation of the cosmetic functional material industry has become the third-generation fermentation technology through the advanced fermentation production technology which can develop only a large amount of useful bioactive substances.

The Korean people use fermented foods such as kimchi, soybean paste and so on, so the concept of fermentation is very familiar and cosmetics using fermented raw materials are used without any objection. In addition, when the fermentation technology is used to develop new materials for the general public, it is known that many harmful ingredients that exhibit completely new effects with existing materials are changed into safe ingredients. Therefore, preference for fermented cosmetics is high, However, it is known that many fermenting materials applied to cosmetics currently use natural fermentation or a fermentation method close to natural aging.

Recently, the trend of domestic fermentation technology has been known to use natural fermentation or a fermentation method close to natural fermentation. Natural fermentation refers to the use of natural raw materials such as water, wind, earth, Although it takes a long time to obtain a variety of substances, it is possible to obtain effects such as moisturizing, antioxidation, wrinkle improvement, cell activity and skin texture improvement. However, natural fermentation or natural fermentation Fermentation is difficult to obtain fermentation products of a certain quality per production batch, and it is also difficult to control quality.

Korea's fermented cosmetics market has been growing at a high rate of 40% over the past three to four years. It is expected to reach about 450 billion won in 2012, about 650 billion won in 2013, and about 1 trillion won in 2014 Have

The fermentation product can be classified into fermentation filtrate and fermentation lysate. The fermentation filtrate utilizes a supernatant in which microorganisms have been removed through a filtration device after the completion of fermentation, and fermentation broth After the fermentation is complete, microorganisms are killed and used by utilizing heat, pH, high pressure of enzyme (Lysozyme) and so on.

In general, fermentation products used in cosmetics are fermented filtrate (fermentation filtrate), which is relatively simple and cost-effective compared to fermented lysate. On the other hand, the fermentation broth is effective in the fermentation of fermented microorganisms in terms of the efficacy of the raw materials, but it is not used frequently because of the additional processing hassles and additional cost problems.

Collagen, which accounts for 70 to 80% of the extracellular matrix, is rapidly degraded as the age increases and becomes closely related to wrinkle formation. Collagen, elastin, Proteoglycans, glucosaminoglycan, laminin, fibronectin, etc. are oxidized and lose their function, causing the skin to lose its elasticity and become wrinkled excessively and become senile skin. Goes. There are collagen fibers, collagen fibers, reticular fibers, elastic fibers (elastic fibers, elastic fibers) in the connective tissue of the extracellular matrix. Collagen, which accounts for 70% Are mostly formed in fibroblasts of the skin. As you get older, the collagen content in the connective tissue of the skin decreases, which is due to degradation of collagen synthesis and promotion of degradation. Therefore, degradation of collagen biosynthesis and promotion of collagen degradation are the major causes of wrinkles in the skin. The collagen biosynthesis process is regulated by many factors involved in transcription level and post-translation level, and collagen degradation is triggered by collagenase degrading collagen by ultraviolet rays Promotion of matrix metalloproteases (MMP), such as collagenase, promotes collagen degradation and reduces collagen content. In addition, as the collagen's deformation accelerates due to the external environment, the skin becomes wrinkled and deepened. As a result, skin aging is caused by non-homogenization of cells, loss of elastin, destruction of collagen, reduction and delay of collagen synthesis, and the like. Therefore, the skin aging phenomenon occurs in the epidermis of the skin but can be seen as a phenomenon that occurs in the dermis.

Accordingly, the present inventor has made a careful effort to develop a fermentation product derived from a natural material exhibiting an effect of preventing skin aging and improving skin wrinkles. As a result, the present inventors have found that a mixture of ice plant and collagen, a succulent plant of the genus Mesembryanthemum , Fermented product fermented by a microorganism or lactobacillus microorganism and a fermentation broth thereof exhibit an excellent anti-aging effect on skin and an effect of improving skin wrinkles, thus completing the present invention.

Accordingly, it is an object of the invention is to provide a Bacillus (Bacillus) or a microorganism of the genus Lactobacillus (Lactobacillus) mixture of ice plant and collagen fermentation using a microorganism of the genus fermentation.

Another object of the present invention is to provide a mixed fermented product characterized in that the mixed fermented product is a fermentation broth.

Another object of the present invention is to provide a mixed fermented product characterized in that the mixed fermented product exhibits an effect of preventing skin aging and improving skin wrinkles.

Another object of the present invention is to provide a method for producing a mixed fermented product of ice plant and collagen comprising the steps of:

(1) preparing an ice plant extract; And

(2) mixing the ice plant extract with collagen, and then inoculating the microorganism of the genus Bacillus or the microorganism of the genus Lactobacillus to perform fermentation.

Another object of the present invention is to provide a method for producing a mixed fermentation product, which further comprises a step of dissolving the fermentation product after the step (2).

Another object of the present invention is to provide a cosmetic composition comprising the mixed fermented product as an active ingredient.

Another object of the present invention is to provide a cosmetic composition which is characterized in that the cosmetic composition is for preventing skin aging and improving skin wrinkles.

According to an aspect of the present invention The present invention provides a Bacillus (Bacillus) or a microorganism of the genus Lactobacillus (Lactobacillus) mixture of ice plant and collagen fermentation using a microorganism of the genus fermentation.

In the present invention, the ice plant is a succulent plant of the genus Mesembryanthemum having a low germination rate and a low growth rate. Origin is Namib Desert, South Africa. The main distribution areas are South Africa, Eastern and Western, Northwest, Southwest Australia, and the arid western part of the Americas. Transparent crystals (bladder cells) on the surface contain a variety of minerals and components useful for the human body, such as inositols and beta-carotene. Bladder cells of stem and leaf have the same shape as ice crystals and are called 'ice plant'. Especially, since the ice plant contains a lot of pinitol which lowers the blood sugar level and myoinositol which suppresses the neutral fat, when the leaves and the stem are reproduced or used as the juice, the healing food roll suitable for the diabetic patients receives the spotlight have. Especially, the plants that grow in the desert have a lot of moisturizing ingredients that can protect the inside of plants to adapt to the water-deficient environment. In the cosmetics industry, they take advantage of this and utilize them as various moisturizing and antioxidant materials Ice plant is a plant that grows in the desert and contains various kinds of moisturizing ingredients inside the plant.

In the present invention, the microorganism of the genus Bacillus is B. subtilis, B. cereus, B. thuringiensis 97-27, B. weihenstephanensis, And B. amyloliquefaciens. ≪ Desc / Clms Page number 7 >

In the present invention, the Lactobacillus sp. Microorganism is selected from the group consisting of Lactobacillus acidophilus, Lactobacillus fermentum, Bifidobacterium bifidum Lactobacillus casei, Lactobacillus reuteri, Lactobacillus bifidum, Lactobacillus gasseri, Lactobacillus plantarum, Lactobacillus johnsonii, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus paracasei, Lactobacillus paracasei, Bacillus crispatus, and the like.

In the present invention, the mixed fermented product may be a fermented seaweed.

The fermentation product can be classified into fermentation filtrate and fermentation lysate. The fermentation filtrate utilizes a supernatant in which microorganisms have been removed through a filtration device after the completion of fermentation, and fermentation broth Means to kill microbes after fermentation by using heat, pH, high pressure of enzyme (Lysozyme) and so on.

In the present invention, the mixed fermented product is characterized by exhibiting an effect of preventing skin aging and improving skin wrinkles.

According to another aspect of the present invention, there is provided a method for producing a mixed fermented product of an ice plant and collagen comprising the steps of:

(1) preparing an ice plant extract; And

(2) mixing the ice plant extract with collagen, and then inoculating the microorganism of the genus Bacillus or the microorganism of the genus Lactobacillus to perform fermentation.

In the present invention, the ice plant extract may be extracted with any one solvent selected from the group consisting of water, alcohols having 1 to 6 carbon atoms, hexane, ethyl acetate, and mixed solvents thereof. The extract of the present invention is preferably extracted with a solvent selected from water, an alcohol having 1 to 6 carbon atoms and a mixed solvent thereof, or a lower alcohol aqueous solution having 1 to 6 carbon atoms. The concentration of the aqueous alcohol solution may be 10 to 100% (v / v), preferably 70 to 100% (v / v). The extraction solvent of the present invention is most preferably an ethanol aqueous solution of 95% (v / v).

The method of extracting an ice plant according to the present invention can be used without limitation as long as it is known in the art, and examples thereof include, but are not limited to, cold-pressing, hot water extraction, ultrasonic extraction, Preferably, it may be hot water extraction.

The extract prepared by the above method is collected by filtration, and the extract is re-extracted for 4 hours under reflux and cooling conditions by addition of an extraction solvent such as water of 4 to 7 times the weight of the raw material to the residue, filtered and mixed with the previous filtrate The extraction efficiency can be increased. The extract of the present invention is not limited to the number of times of extraction.

After the extraction as described above, the combined filtrate is concentrated under reduced pressure at 40 to 60 ° C to remove the solvent remaining in the extract, and then azeotropically concentrated with water 25 to 50 times the weight of the concentrate under reduced pressure for 2 to 3 times, And the mixture is further homogenized. The final extract may be further purified, or may be dried by freeze-drying or the like to remove the solvent to solidify it, or may be prepared in powder form.

After the ice plant extract is mixed with collagen, the microorganism of the genus Bacillus or the microorganism of the genus Lactobacillus is inoculated and fermented.

The mixing ratio of the ice plant extract and collagen is not particularly limited and may be mixed in a ratio of 1: 0.001 to 5.

The concentration of the microorganism in the ice plant extract and collagen mixture, the fermentation temperature, and the fermentation time may be the same as those known in the art.

The present invention further provides a method for producing a mixed fermentation product, which further comprises a step of dissolving the fermentation product after the step (2).

The fermentation broth of the mixed fermented product can be produced by the above method.

In order to achieve another object of the present invention, there is provided a cosmetic composition comprising the mixed fermented product as an active ingredient.

In the present invention, the cosmetic composition may be characterized by exhibiting an effect of preventing skin aging or improving skin wrinkles.

The cosmetic composition may be applied to a gel type, a skin type, a cream type, an ointment type, and the like, but is not limited thereto. The above composition can be suitably prepared by known methods, adding appropriate conventional softening agents, emulsifying agents, thickening agents or other materials known in the art depending on the type thereof.

The gel-type composition may be prepared by adding a softening agent such as trimethylolpropane, polyethylene glycol or glycerin, a solvent such as propylene glycol, ethanol, isocetyl alcohol, or the like.

The skin-type composition may contain at least one of fatty alcohols such as stearyl alcohol, myristyl alcohol, behenyl alcohol, arachidyl alcohol, isostearyl alcohol and isocetyl alcohol, butylene glycol, 2-sodium, xanthan gum, dimethicone, polyethylene glycol-60 hydrogeneate castor oil, polysorbate 60 and purified water.

The cream-type composition may be formulated by dissolving or dispersing the above-mentioned cream type composition in an aqueous medium such as alcohols such as stearyl alcohol, myristyl alcohol, behenyl alcohol, arachidyl alcohol, isostearyl alcohol, isocetyl alcohol and the like, lecithin, phosphatidylcholine, phosphatidylethanolamine, Emulsifiers such as glyceryl stearate, sorbitan palmitate, and sorbitan stearate; natural fats or oils such as avocado oil, apricot oil, barba water oil, glass fat oil and camellia oil; Propylene glycol and the like, purified water, and the like.

The ointment type composition may be prepared by adding a softening agent, an emulsifying agent, and a wax such as microcrystalline wax, paraffin, ceresin, wax, spermaceti, vaseline or the like.

The cosmetic composition of the present invention may further contain at least one ingredient showing the same or similar functions in addition to the effective ingredient. Non-limiting examples include, but are not limited to, one or more selected from the group consisting of vitamin C, retinoic acid, TGF, protein from animal placenta, betulinic acid and chlorella extract. The cosmetic composition of the present invention may further contain an excipient including a fluorescent substance, a fungicide, a hygroscopic substance, a moisturizing agent, a fragrance, a fragrance carrier, a protein, a solubilizer, a sugar derivative, a sunscreen, a vitamin, .

The ice plant and collagen-based fermented product and fermented seafood according to the present invention exhibit remarkably improved skin aging-preventing effect and skin wrinkle-improving effect as compared with non-fermented product, and thus can be very usefully used for manufacturing functional cosmetics.

Fig. 1 is a table showing the activity of inhibiting elastase according to the extraction condition of the ice plant.
FIG. This is a comparison of MMP-1 activity by ice plant extraction conditions.
3 is a graph showing a comparison of microbial growth during fermentation by lactic acid bacteria.
FIG. 4 is a graph showing the activity of inhibiting elastase upon fermentation by lactic acid bacteria.
FIG. 5 is a graph showing MMP-1 activity evaluation by ice plant and collagen fermentation products.
FIG. 6 is a photograph of a skin primary stimulation test of an ice plant and a collagen fermented product. FIG.

Hereinafter, preferred embodiments of the present invention will be described in detail. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the accompanying drawings. Throughout the specification, when an element is referred to as " including " an element, it means that it can include other elements, not excluding other elements, unless specifically stated otherwise.

Example

Example 1: Identification of extraction conditions of ice plant

The antioxidant activity in plants is mainly represented by phenolic components such as flavonoids, polyphenols, tannins, and phenolic terpenes. The extraction yield of each phenolic component varies depending on the plant. However, when the plant is heat- The component becomes free and increases the activity. Therefore, it has been reported that the extracts of hot water have higher antioxidant activity and tyrosinase inhibitory activity than other organic solvent extraction methods, and that they have excellent ability to inhibit pancreatic lipase, a-glucosidase and NO production by extraction with organic solvents The fermentation was carried out using hot water extraction and organic solvent extraction, and the efficacy was evaluated to select the most effective extraction method.

The ice plant was subjected to hot water extraction, 70% ethanol extraction, and simultaneous extraction to perform the combined fermentation of the collagen-ice plant. elastase inhibitory activity and MMP-1 activity, the process was established by the simultaneous extraction of the ice plant extraction method. (Figures 1 and 2)

Example 2: Ice plant collagen fermentation and fermentation product identification

Minimum culture culture experiments were carried out to obtain fermentation conditions with optimum antioxidative effect by using ice plant extract and polymer collagen (Mw 100,000 ~ 300,000) obtained from Bacillus genus and Lactobacillus genus in different fermentation conditions.

Lactobacillus acidophilus , Lactobacillus plantanum, Lactobacillus rhamnosus, Lactobacillus paracasei and Bacillus subtilis were used as the strains.

In the specific culture method, the above four strains were purely cultured in a selective medium, and inoculated with 1% of the culture medium in which the number of bacteria of each strain was about 10 ⁸ to 10 ⁹ cfu / ml, and cultured in a 37 incubator for 72 hours . The culture medium used was 5 g of peptone, 2.5 g of sodium acetate trihydrate, 0.5 ml of magnesium sulfate heptahydrate, 0.5 ml of manganese sulfate tetrahydrate, 5 ml of tween 80, 1 g of diammonium citrate, 1 g of dipotassium phosphate phosphate 1 g, and distilled water 480 ml, were mixed with 15 g of collagen.

After the cultivation was completed, the culture broth was centrifuged at 4000 rpm for 10 minutes to separate the lactic acid bacteria and the culture broth, and the separated culture broth was confirmed to have low molecular weight collagen by SDS-PAGE.

As a result of 5 kinds of lactic acid bacteria tested, L. paracasei and L. rhamonosus were the most effective in the growth of collagen - ice plant by lactic acid bacteria among 5 lactic acid bacteria, and L.paracasei Were found to be most suitable for fermentation. (Figs. 3 and 4)

Example 3: Preparation of fermented seafood and inhibition of MMP-1 (Matrix Metalloproteinase-1) activity

After fermentation, the fermentation broth was fermented to prepare fermentation broth. Lysozyme treatment and microfluidizer treatment were applied to the fermentation broth, and the broth was filtered with 0.22um filter to completely remove residual microorganisms. Seafood was prepared. The inhibitory effect of collagenase (MMP-1) was then tested to confirm the anti-aging effect of the lysate.

Human dermal fibroblast (HDF) cells were cultured in DMEM (Dulbecco's modified Eagle's media, Sigma) supplemented with 10% fetal bovine serum (Sigma) at 37, 5% CO2. Cells were cultured on a 24-well plate at a concentration of 2 X 105 cells / well. After confirming cell attachment, only the solvent was added to the control group without any treatment. In the dish, 20 kinds of retinol and 10 ug / ml. < / RTI > The test material was added to each dish and cultured for 2 days. After 2 days, the medium was harvested and the inhibitory activity of MMP-1 was measured by Matrix Metalloproteinase-1 (MMP-1), Human, biotrak, ELISA kit (GE healthcare RPN2610)

After fermentation (fermentation filtrate) and fermentation broth were treated at different concentrations, the inhibitory effect of MMP-1 was measured. From these results, it can be shown that fermented seaweed is more useful than simple fermentation filtrate when applied UV as a factor causing MMP-1 production in skin, thus inhibiting MMP-1 production and suppressing skin wrinkles .

Example  4: Ice plant, collagen Fermentation  Clinical trial approval and subject recruitment

 1) Clinical trial IRB application and approval

  (1040647-201704-HR-008-02) approved by the Bioethics Committee of Daejeon University to conduct whitening and moisturizing clinical trials. The experimental group consisted of Lactobacillus fermented product (with / without ice), Saccharomyces fermented product (commercial product) and bass cream group, and the subject's face was divided into two parts.

  Apply cream at random and apply each cream twice a day in the morning and evening, and apply directly to the face as a visual and instrumental evaluation before the experiment, 2 weeks, 4 weeks, 6 weeks and 8 weeks after application (2022) and relative humidity (4060%) were maintained before and after the experiment, and the subjects were rinsed with lukewarm water using neutral detergent. For the measurement in the state of recovery of the skin, take a stable measurement for 1020 minutes, and then perform a skin test. The camera is used to fix the angle so that the test site is in the same field of view. In addition, the subject and the camera position will be fixed so that the distance between the test site and the lens will be the same every time.

  Clinical Trials IRB was completed and approved based on the above information, and entered the clinical trial participants recruitment.

   2) Recruitment of clinical trial subjects

  Subjects were recruited from the 2050s who live in Daejeon Metropolitan City and Chungcheongnam-do. According to the guidelines for evaluating the effectiveness of functional cosmetics announced by the Korea Food & Drug Administration in 2015, the minimum specimen size to be recruited when 20 experimental groups and 20 control groups were set (20 groups of 4 = 80) (= 40 persons). In other words, 50 people will be recruited considering the dropout rate of 20%.

The subjects of this study were selected as follows.

        - A person who voluntarily agrees to participate in the study and who has signed and signed the participation agreement

        - A woman aged 20 and 50 years old who is suffering from hyperpigmentation

        - Those who can read and respond to the questionnaire

        - Healthy person who does not have acute and chronic physical illness including skin disease

       The criteria for exclusion of subjects in this study are as follows.

        - Pregnant or lactating women and women who are pregnant

        - Those who use a skin contour containing steroid for more than 1 month for the treatment of skin disease

        - Those who have not passed six months after participating in the same experiment

        - Persons with sensitive, irritable skin

        - Those who have skin abnormalities such as dots, acne, erythema and capillary dilatation on the sample application area.

        - Those who use the same or similar cosmetics and medicines in the study area within 3 months before the start of the study

        - Those who are deemed to be unsuitable for the experiment at the discretion of the other state tester.

       The criteria for dropping out of the study subjects are as follows.

        - Those who have erythema or pruritus during sample application

        - Excessive UV exposure during the experiment

        - Those who are deemed to be unsuitable for the experiment due to the judgment of the state tester.

   3) Skin primary stimulation test

     Test design

     . Test substance: Ice plant Collagen Fermentation substance / Control test substance

     . As a preliminary experiment, a room for six people

     . Conducted to all subjects before experiment

     . Substitute skin under the skin like a face

     . Measures pigmentation, viscosity, pH, humidity, room temperature, etc.

     Test method (48 hr single patch test)

     . Wipe with 70% alcohol and dry

     . A certain amount (after application amount) was loaded into a patch test chamber (IQ Ultra TM)

     . Place the chamber on the upper left underneath of the subject and fix it with a microtape.

     . Remove after 48 hours

     . After 30 minutes and 24 hours after removal of the patches

    Primary skin irritation preliminary test

     . Period: April 26, 2017 - April 29, 2017

     . Target audience: 6 women in their 20s and 30s

     . Experimental group:

        Ice Plant Collagen Fermentation Material

        Positive control substances

        Base cream (negative control)

3) Results:

Visual Observation There were no problems in using 6 persons and it was confirmed that there were no visible problems such as erythema. (Fig. 6)

The preferred embodiments of the present invention have been described in detail above. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Accordingly, the scope of the present invention is defined by the appended claims rather than the foregoing detailed description, and all changes or modifications derived from the meaning, range, and equivalence of the claims are included in the scope of the present invention Should be interpreted.

Claims (7)

Bacillus (Bacillus) or a microorganism of the genus Lactobacillus (Lactobacillus) mixture of ice plant and collagen fermentation using a microorganism of the genus fermentation. The method according to claim 1,
Wherein the mixed fermented product is a fermentation broth. The method according to claim 1,
Wherein the mixed fermented product exhibits an effect of preventing skin aging and improving skin wrinkles. A method for producing a mixed fermented product of an ice plant and collagen comprising the steps of:
(1) preparing an ice plant extract; And
(2) mixing the ice plant extract with collagen, and then inoculating the microorganism of the genus Bacillus or the microorganism of the genus Lactobacillus to perform fermentation. 5. The method of claim 4,
And further comprising the step of dissolving the fermented product after the step (2). A cosmetic composition comprising the mixed fermented product of claim 1 or 2 as an active ingredient. The method according to claim 6,
Wherein the cosmetic composition is for preventing skin aging and improving skin wrinkles.

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