KR101821955B1

KR101821955B1 - Process For Preparation Of Composition For Improving Hair Health

Info

Publication number: KR101821955B1
Application number: KR1020150096174A
Authority: KR
Inventors: 옥민; 윤경훈; 박진철; 박상현; 성혜영; 강진; 박지연; 홍우종; 박성수
Original assignee: (주)엠앤씨생명과학; 옥민
Priority date: 2015-07-06
Filing date: 2015-07-06
Publication date: 2018-01-26
Also published as: KR20170005726A

Abstract

The present invention provides a composition for promoting hair health comprising a fermented yeast cake fermented extract as an active ingredient and a method for producing the same.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a hair growth promoting composition,

The present invention relates to a method for producing a composition for promoting hair health.

Hair is produced from hair follicles (follicles) present below the surface of the skin (Fig. 1). When the hair follicle is formed by the interaction of the epidermis and the dermis, the hair grows from the hair follicle. Below the hair follicle is a dermal papilla.

The hair is about 100,000 pieces, and the whole hair grows about 0.3 ~ 0.4 ㎜ per day. This rapid growth of hair is due to the rapid, vigorous cell division that can not be found in any part of the body in hair matrix cells that correspond to its roots.

Each hair has a different growth period. FIG. 2 shows a hair growth cycle, which can be divided into four stages.

The first stage is the anagen stage where hair grows most actively. Papilla, which is fed from capillary blood vessels, provides nutrients to the hair cells, resulting in smooth cell division and thus new hair.

The second stage is the catagen stage where hair regrowth begins. The follicular part starts to slowly contract and separate from the dermal papilla. It is short in about several weeks. No more neonatal cells are produced and the cell division of the mononuclear cells gradually decreases.

The third stage is the telogen stage where hair growth stops or rests. The mothers are completely separated from the papillae and no more newborns develop. The hair itself is just present in the hair follicle, and the hair easily falls off even in weak stimuli.

The fourth step is an early anagen that starts to produce new hair. At the end of the dormant period, the embryonic cells under cell division divide again to create new hair as they come into contact with the dermal papilla.

This cycle repeats over 3 to 6 years and results in an average of 50 to 200 hairs per day.

Hair loss generally refers to a decrease in the number of hairs in the growing season and an increase in the number of hairs in the retrogressive or resting period, resulting in an abnormally high number of hairs. If a slight disruption occurs in the rapid cell division in the hair follicle, hair growth may be immediately affected and hair loss may occur.

The causes of hair loss include male hormone excess, poor blood circulation, peroxidation, bacteriostatic dysfunction of the scalp, genetic factors, aging, and stress have been discussed, but until now the cause is not clear. Recently, due to nutritional imbalance caused by dietary changes, environmental pollution, and stress caused by the social environment, the number of people suffering from hair loss is increasing, and the age is also lowered and the female hair loss population is also increasing.

Until now, no precise treatment for hair loss has been suggested. Wig, and mastectomy are suggested, but it can not be a fundamental solution to hair loss. Hair dyes, hair growth agents, and the like are being marketed without clarifying the cause and mechanism of alopecia.

It is known that hair growth promotes blood circulation or nutrition, promotes metabolism of hair follicles, inhibits reductase, and reduces fat, but it has not yet had a great effect . Representative hair growth products currently marketed include minoxidil and propecia.

Minoxidil was originally used as an antihypertensive agent, but it was identified as a side effect by patients who used this drug and was used as a hair growth promoter. Here, 2% minoxidil is used as a hair growth agent to which attention is paid. When minoxidil is used alone, it is not absorbed well and it must be used first after applying a skin irritant called 'Tretinoin' (Retin-A), so it is absorbed well and effective. Minoxidil should also be used continuously. In this case, blood pressure may be affected. If it is discontinued, hair roots become weak and fall off.

Propecia (Pinasteride) was originally used as a treatment for benign prostatic hyperplasia, but side effects were observed in patients taking the drug, developed as a treatment for male hair loss. Propecia inhibits the action of 5α-reductase and inhibits the production of DHT to prevent and treat male pattern hair loss. Propecia inhibited the production of DHT, reducing the levels of DHT in the scalp and serum by 60% and 70%, respectively, without affecting testosterone, which is involved in male sexual function. However, taking this medicine may have side effects such as sexual dysfunction.

Others In Korea, Japan, and China, various types of hair growth promoting herbs are known.

Korean Patent Laid-Open Publication No. 10-2001-0100497 discloses a hair growth promoting agent comprising extracts such as mugwort, pseudo-medicinal plant, Seokjanghwang, Angelica gigas, Hwanggi, Seokjangbok, Ginkgo biloba, Mulberry leaf, etc., and Patent Document 10-2004-0001683 'Natural blooms using extracts such as pine needle, ginkgo leaf, mulberry root, and gingko mushroom' have been introduced, but their efficacy is minimal and there is no scientific basis.

The above-mentioned hair remedies have been known to stimulate blood circulation of the scalp to prevent hair loss or promote hair growth, but the effect has not been proved yet, the effect is extremely low, and in some cases, side effects such as scalp hypersensitivity have appeared.

Korean Patent Publication No. 10-2001-0100497 Korean Patent Publication No. 10-2004-0001683

Kim, Gyu-Han, 'Effect of minoxidil on proliferation and apoptosis in human breast-hood' (Journal of Dermatological Science 34, 91-98, 2004)

The present invention solves the above problems of the prior art and provides a hair health promoting composition comprising a yeast cake fermented extract capable of stimulating hair dermal papilla cells to prevent hair loss without promoting toxicity, And a method thereof.

One aspect of the present invention is a method for producing a kale extract, comprising: adding kale to distilled water and heating at 60 to 100 ° C for 1 to 3 hours to obtain a kale extract;

delete

Step of obtaining a kale fermentation extract by inoculating the yeast with Saccharomyces cerevisiae yeast in the kale extract and fermenting at 10 to 50 ° C for 1 to 3 days;

And a method for producing a composition for promoting hair health.

delete

According to one embodiment of the present invention, it is possible to provide a composition for promoting hair health and a method for producing the composition, which can improve hair health by preventing hair loss or promoting hair growth without causing toxicity problems. Accordingly, the composition according to the present invention can exert excellent effects on safety and hair health.

Fig. 1 shows the structure of a general hair.
Fig. 2 shows a hair growth cycle.
Fig. 3 is a graph showing the results of measurement of the absorbance of minoxidil, hydrothermally extract (kale), and yeast kale fermentation extract (kale Y1).
Fig. 4 is a graph showing the protein expression effect of the untreated group, minoxidil, hydrothermal extract (kale), and yeast kale fermentation extract (Y-kale).
FIG. 5 shows a microscopic photograph (100X magnification) for confirming the cell survival / proliferation degree of the untreated group, minoxidil, hydrothermal extract (kale) and yeast kale fermentation extract (Y-kale).
FIG. 6 is a graph showing the results of measurement of protein Bax expression of untreated group, minoxidil, hydrothermal extract (kale), and yeast kale fermentation extract (Y-chain).

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will now be described in detail with reference to the accompanying drawings.

However, these descriptions are provided only to illustrate the present invention, and the scope of the present invention is not limited by these exemplary explanations.

The present invention provides a composition for promoting hair health comprising a fermented yeast cake fermented extract as an active ingredient. Here, the yeast-cake fermentation extract is obtained by fermenting kale by inoculating yeast into the kale extract.

As the yeast, Saccharomyces cerevisiae MAB Y1 (accession number: KCTC 11386BP) is preferably used.

The formulation of the composition is not particularly limited and may be selected from the group consisting of a shampoo, a rinse, a tonic, an essence, a lotion, a conditioner, a foam, a gel, a setting lotion, a hair stick, a blow, a zymo coat, a wave agent, Mousse, aerosol, pomade, powder, soap, pack, ointment or cream formulation.

The method of making the composition

Adding kale to distilled water and heating to obtain a kale extract,

Obtaining a kale fermentation extract by inoculating Saccharomyces cerevisiae MAB Y1 (accession number: KCTC 11386BP) as yeast into the kale extract by fermentation;

Concentrating the kale fermentation extract and lyophilizing

.

Here, kale is the highest-beta-carotene vegetable among green-yellow vegetables. Kale contains vitamins, minerals, amino acids, fats, proteins, and fiber. It has a blood-forming blood-forming effect and promotes metabolism.

Specifically, kale is added to distilled water, and then the kale extract is prepared by heating at 60 to 100 ° C for 1 to 3 hours. The obtained extract is subjected to high pressure steam sterilization at 100 to 140 ° C.

The primary kale extract is inoculated with the sugar and the yeast, and the fermented extract is fermented at 10 to 50 ° C for 1 to 3 days to produce a primary kale fermentation extract. The concentrated fermented extract solution is used or concentrated to produce a concentrate.

The fermented product of the primary kale is separated into a residue and an extract. The solvent is then added to the residue to extract at 60 to 100 ° C. The primary kale fermentation extract separated from the primary kale fermentation extract and the secondary fermentation extract are combined The obtained fermentation extract is concentrated to produce a concentrate.

The obtained fermented extract concentrate is preliminarily frozen at -90 to -50 ° C for 6 to 10 hours and then lyophilized to finally produce an extract.

In the hair, the dermal papilla cells play an important role in regulating the hair growth cycle and supplying the nutrition by participating in the hair growth and growth. As shown in FIG. 1, the dermal papilla cells form cells of hair follicles together with dermal cells, which absorb nutrients from capillaries and interact with dermal cells to form and grow hair.

Studies have shown that activating signaling of extracellular signal-regulated kinases (ERKs) and Akt (protein kinase B), and increasing Bcl-2 expression and decreasing Bax expression, (Kim, Gyu - Han, Effect of minoxidil on proliferation and apoptosis in human dermal papilla cells).

Here, Bcl-2 acts as an antioxidant in mitochondrial nuclear membrane and endoplasmic reticulum, inhibits lipid peroxidation, inhibits active noxious oxygen production, blocks Ca2 + influx into the nucleus and mitochondria, and activates caspase And inhibit the outflow of cytochrome C, and so on. Thus, Bcl-2 functions to suppress apoptosis.

Bax, on the other hand, is known to inhibit Bcl-2 function and induce apoptosis.

When the concentration of active oxygen is low, the extracellular signal-regulated kinases (ERK) protein, which is a typical signal transducer involved in cell survival and proliferation, is activated.

The composition according to one embodiment of the present invention contributes to the promotion of hair health by reducing the protein involved in cell death and helping to produce proteins involved in the survival / proliferation of the papilla cells, do. Specifically, it helps prevent hair growth and hair loss.

Preparation Example: Preparation of Yeast Kale Fermented Extract

1. Preparation of primary kale extract

160 g of kale (Oceana, Gyeongnam Hamman) was crushed and put into a flask containing 400 ml of distilled water and extracted at 80 DEG C for 2 hours. The obtained extract was autoclaved at 121 占 폚 for 15 minutes under high pressure.

2. Preparation of primary kale fermented extract

2.5% by weight of sucrose, 5% by weight of saccharomyces cerevisiae MAB Y1 and KCTC 11386BP were added to the primary kale extract, and the mixture was inoculated at 30 ° C for 1 day to give a fermented extract of primary kale Respectively.

3. Preparation of fermented extract of secondary kale

The fermented extract of the primary kale was separated into a residue and an extract, and 400 ml of ethanol was added to the residue, followed by extraction at 80 ° C for 2 hours.

4. Enrichment

The fermented extract of the primary kale isolated from the primary kale fermentation extract and the secondary fermentation extract were mixed, and then the total 800 ml of the fermented extract was concentrated to finally produce 30 ml of the concentrate.

5. Freeze-dried

The fermented extract concentrate (30 ml) was preliminarily frozen at -70 ° C for more than 8 hours and then lyophilized to finally produce an extract. Distilled water was added to the extract to obtain a stock solution at a concentration of 1000 mg / ml.

Experimental Example

[Cell Culture of Experimental Subject]

Human dermal papilla cells (HHDPC) were purchased from ScienCell (Diego, CA, USA).

Mesenchymal stem cell medium (MSCM, ScienCell # 7501) supplemented with 5% FBS (fetal bovine serum) and 100 U / ml penicillin / streptomycin and MSCG (mesenchymal stem cell growth supplement; ScienCell # At 37 ° C and 5% CO ₂ .

Experimental Example 1: Stability test

MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide) assay

MTT assay was performed to examine the effects of minoxidil, hydrocele extract and yeast kale fermentation extract on cell viability. MTT assay is a test using mitochondrial ability to reduce the yellow water soluble substrate MTT tetrazolium to blue-violet non-water soluble MTT formazan. The MTT assay measures the absorbance of MTT formazan, and the measured absorbance is alive and metabolized It is well known that it reflects the concentration of cells.

First, human dermal papilla cells (HHDPC) were dispensed into 96-well plates at a density of 1.0 × 10 ⁴ cells / well using ScienCell (Diego, CA, USA). The human dermal papilla cells were treated with minoxidil, hydrothermal extract and yeast kale fermentation extracts at 1, 5, 10, 25, 50 and 100 μg / mL, respectively. After 24 hours, the treated dermal papilla cells were incubated in MTT (1 mg / ml) solution for 4 hours. It was then dissolved in 0.04 N HCl / isopropanol and the absorbance at 570 nm was measured with a spectrophotometer.

The absorbance measurement results are shown in Fig. In FIG. 3, the cell viability is the MTT absorbance value of the sample treated as divided by the MTT absorbance value of the sample that was not treated, and then multiplied by 100, in%.

In the case of minoxidil, the cell survival rate was decreased by about 7% at the concentration of 5 μg / mL (about 93%), and the cell survival rate was decreased as the concentration was continuously increased. 56%, and only about 13% at the concentration of 100 μg / mL. As a result, it was observed that the toxicity of minoxidil was remarkably increased when the concentration exceeded a certain concentration. In contrast, the hydrothermal kale extract (Kale) and the yeast kale fermentation extract (Kale Y1) were found to be safe without cytotoxicity regardless of the concentration.

Experimental Example 2: Analysis of protein expression effect

Effect of Protein Expression Enhancement in Cell Survival / Proliferation of Yeast Kale Fermentation Extracts (Protein Isolation and Western Blot Analysis)

(2) human dermal papilla cells were treated with 25 μg / ml minoxidil, (3) human dermal papilla cells were treated with 5, 10 and 25 μg / ml of hydrothermal kale extract (4) human dermal papilla cells treated with 5, 10, and 25 μg / mL of yeast-cake fermentation extract. Thus, a total of 8 samples were prepared.

The samples were subjected to western blotting in order to measure the expression of active ERK-1,2 protein.

Cells were treated with lysis buffer to prepare cell extracts, and 10% SDS-PAGE was performed on the proteins of the cell extracts. The electrophoretic protein bands were transferred to a nitrocellulose membrane and treated with 5% skimmed milk powder to prevent nonspecific binding. Then, the primary antibody of ERK1 / 2, Phospho-ERK1 / 2 protein (monoclonal rabbit anti- Tech., # 4695, # 9101) was reacted with 1,000-fold dilution and reacted with secondary antibody of goat anti-rabbit horseradish peroxidase (1: 10000, Jackson ImmunoReasearch Lab.) For 1 hour with stirring. ERK 1/2 and pERK 1/2 proteins in the nitrocellulose membrane were detected with a chemiluminescent reagent kit (Detroit R & D Inc.) and photographed with an X-ray film (Konica Co.).

The measurement results are shown in Fig.

As shown in FIG. 4, the fermented yeast cake fermentation extract significantly increased the expression of phospho-ERK in comparison with the untreated group, minoxidil and hot water extraction kale. Phosphor-ERK is an active form of ERK-1 and 2 proteins involved in the survival / proliferation of dermal papilla cells. Therefore, it can be seen that the yeast kale fermentation extract increases the survival / proliferation effect of the dermal papilla cells.

Microscope observation and photograph

(2) human dermal papilla cells treated with 25 μg / ml minoxidil, (3) human dermal papilla cells treated with 25 μg / ml of hydrothermal kale extract, (4) human dermal papilla cells treated with 25 μg / mL of yeast-kale fermentation extract. Four samples were thus prepared.

In order to confirm the degree of cell survival / proliferation in the sample, the sample was observed with a microscope (Leica Microsystems, Swizerland) at a magnification of 100X and images were obtained.

The photographed image is shown in Fig. As shown, the yeast kale fermentation extract showed the highest cell survival / proliferation even under a microscope.

Experimental Example 3: Analysis of inhibitory effect on protein expression

Inhibitory Effect of Yeast Kale Fermentation Extracts on Protein Expression in Cell Death (Protein Isolation and Western Blot Analysis)

Western blots were performed on the samples to determine Bax protein expression.

Cells were treated with lysis buffer to prepare cell extracts, and 10% SDS-PAGE was performed on the proteins of the cell extracts. The electrophoretic protein bands were transferred to a nitrocellulose membrane and treated with 5% skim milk powder to prevent nonspecific binding. Then, the primary antibody of ERK1 / 2, Phospho-ERK1 / 2 protein (monoclonal rabbit anti- Tech., # 4695, # 9101) was reacted with 1,000-fold dilution and reacted with secondary antibody of goat anti-rabbit horseradish peroxidase (1: 10000, Jackson ImmunoReasearch Lab.) For 1 hour with stirring. The Bax protein in the nitrocellulose membrane was detected with a chemiluminescent reagent kit (Detroit R & D Inc.) and photographed with an X-ray film (Konica Co.).

The measurement results are shown in Fig.

As shown in FIG. 6, the fermentation extract of yeast kale was found to significantly reduce the expression of protein Bax, which is involved in apoptosis, in comparison with minoxidil.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention. The present invention is not limited to the drawings.

Claims

Adding kale to distilled water, heating at 60 to 100 ° C for 1 to 3 hours to obtain a kale extract,
Step of obtaining a kale fermentation extract by inoculating the yeast with Saccharomyces cerevisiae yeast in the kale extract and fermenting at 10 to 50 ° C for 1 to 3 days;
Wherein the hair growth promoting composition is a hair growth promoting composition.

delete

The method according to claim 1,
Wherein the yeast is Saccharomyces cerevisiae MAB Y1 (accession number: KCTC 11386BP).

The method according to claim 1,
The formulations may be in the form of a shampoo, a rinse, a tonic, an essence, a lotion, a conditioner, a foam, a gel, a setting lotion, a hair stick, a blow, a zymo coat, a wave, a hair dye, Powder, soap, pack, ointment or cream formulation.

The method according to claim 1,
And concentrating and freeze-drying the fermented kale extract.

6. The method of claim 5,
After the step of obtaining the kale fermentation extract, before the step of concentrating and lyophilizing the kale fermentation extract,
Separating the kale fermentation extract into a residue and an extract, adding ethanol to the residue to obtain a fermentation extract of the secondary kale,
Mixing the kale fermentation extract separated from the kale fermentation extract and the second kale fermentation extract
&Lt; / RTI > wherein the composition further comprises a surfactant.