KR101902425B1 - Agent for improving regeneration comprising Gardenia jasminodes extracts - Google Patents

Abstract

The present invention discloses a skin regeneration promoting agent and a hairpin activity promoting agent comprising a gardenia extract as an active ingredient.

Description

{Agent for improving regeneration comprising Gardenia jasminodes extracts}

The present invention relates to a skin regeneration and hair growth promoting agent comprising a gardenia extract.

The hair loss can be classified into resting hair loss and growth-type hair loss according to the growth cycle. The resting hair loss hair loss includes male pattern hair loss caused by androgen, female postpartum hair loss or hair loss due to endocrine diseases, Hair loss, scarring hair loss, or hair loss due to head tinea. It is also known that hair loss is caused by local infections, endocrine disorders, genetic factors and autoimmunity.

Currently, representative anti-hair-loss agents include propecia and minoxidil approved by the US FDA. These drugs have a clinically obvious effect, but they also show adverse effects such as sexual dysfunction and skin irritation, and women can only use it in an unusable or extremely limited amount. Therefore, there is a growing interest in natural product preparations that can be used safely without any gender limitation, while still having a disadvantage as described above.

Fathke et al., "Wnt signaling induces epithelial differentiation during cutaneous wound healing ", BMC Cell Biology 2006, 7: 4

The present invention is to provide a skin regeneration promoting agent having an excellent skin regeneration promoting effect and a hair growth promoting agent having an excellent promoting activity of the hair regrowth activity.

One aspect of the present invention provides a skin regeneration promoting agent and a hair follicle activity promoting agent comprising a gardenia extract as an active ingredient.

Another aspect of the present invention provides a cosmetic composition and a pharmaceutical composition comprising the accelerator.

The formulation according to the present invention promotes the transmission of the Wnt / beta-catenin signal by containing the gardenia extract as an active ingredient, thereby exhibiting excellent skin regeneration effect, specifically excellent skin wound healing and scar reduction effect have. In addition, the agent not only promotes the movement of the hair / beta-catenin signal, but also activates the stem cells of the dermal papilla cells and promotes the proliferation of the dermal papilla cells of the hair follicles, .

FIG. 1 is a graph showing that Gardenia extract promotes Wnt / beta-catenin signaling of stem cells.
FIG. 2 is a graph showing that the gardenia extract maintains and activates stem cell lines of the papilla cell line.
FIG. 3 is a graph showing the effect of the gardenia extract on promoting hair follicle dermal papilla cell proliferation.
4 is a graph showing a visual evaluation result showing the hair growth promoting effect of the gardenia extract.
FIG. 5 is a hair weight result graph showing the hair growth promoting effect of the gardenia extract.
Fig. 6 is a photograph showing the results of hair growth after applying the gardenia extract.

As used herein, the term "skin " refers to a tissue covering an animal's body surface, and is a broad concept including not only a tissue covering a body surface such as a face or a body but also a scalp. In the present specification, the term " hair " is a generic term for hair and hair and may also be described as hair. The hair includes both human and animal hair. As used herein, " promoting wool " means not only promoting the creation of new hair, but also allowing existing hair to grow coarser and healthier. As used herein, the term " extract "is a broad concept that encompasses all of the substances obtained by extracting components of natural products, regardless of the type of extraction method, extraction solvent, extracted component or extract.

In each tissue of the human body, there are adult stem cells that produce the cells constituting the tissue. Cells that produce skin or hair are also derived from stem cells present in the skin or hair follicles.

Stem cells generally refer to pluripotent cells that can develop into any tissue. It has a self-renewal ability to produce itself by repeated division and a differentiation ability to differentiate into cells having specific functions depending on the environment. These stem cells can be roughly divided into embryonic stem cells and adult stem cells.

Embryonic stem cells are derived from fertilized eggs produced by fertilization of spermatozoa, male reproductive cells, and female reproductive cells. It has the ability to differentiate into various types of cells and is also called a fully differentiating stem cell. Embryonic stem cells are capable of mass proliferation and differentiation into almost all body cells, and have immunity rejection, which is advantageous in that they can be transplanted into other species as well as others. However, since it is difficult to control the differentiation, it may become a cancer cell. Therefore, it requires precise technology and it may be ethically problematic because embryo is destroyed.

Adult stem cells generally refer to cells that, when needed, differentiate into cells of a particular tissue, and usually only a small amount is present in each tissue of the body. That is, the adult stem cells of the bone marrow cells are defined as hematopoietic cells, the skin adult stem cells as the skin, and the adult stem cells of the olfactory nerve cells as the olfactory neurons. These adult stem cells are not likely to become cancer cells due to their stable differentiation, and unlike embryonic stem cells, they do not destroy embryos and thus do not have ethical problems.

Follicular stem cells are adult stem cells that are present in hair follicles and make up cells that constitute skin and hair. They are follicular stem cells, which can be derived from epithelial cells, also called bulge stem cells, stem cells derived from mesenchymal stem cells And melanin-forming stem cells derived from neural stem cells.

Hereinafter, the present invention will be described in detail.

One aspect of the present invention provides a skin regeneration promoter comprising a gardenia extract as an active ingredient.

Wnt / beta-catenin (Wnt / beta-catenin) signaling is a signaling pathway associated with the development of life, and is known to act on skin formation and hair follicle development. In addition, it is known that when a skin injury occurs, a winter signaling is essentially expressed in the repair of skin tissue, thereby promoting the differentiation of epithelial cells (Fathke et al., "Wnt signaling induces epithelial differentiation during cutaneous wound healing" 2006, 7: 4). Therefore, stimulation of the winter signal transmission promotes differentiation of epithelial cells, thereby promoting skin regeneration and further promoting healing of skin wounds. The gardenia extract promotes the Wnt / beta-catenin signaling of stem cells. Therefore, the preparation containing the same can promote skin regeneration, and can specifically promote the healing of skin wounds.

Another aspect of the present invention provides a hair follicle activity promoting agent comprising a gardenia extract as an active ingredient.

Activation of the stem cells of the dermal papilla cells promotes the activity of the dermal papilla cells and can promote hair growth. Gardenia extract not only activates the stem cells of the dermal papilla cells but also promotes the proliferation of the dermal papillary cells. Therefore, the formulation containing the gardenia extract promotes the activity of the dermal papilla to promote hair growth, and furthermore, it can have the effect of preventing hair loss and promoting wool.

On the other hand, hair repeats growth and elimination through the period of growing period - period of regenerating - period of resting period. In the growth phase, new hair is formed as the hair follicle stem cells differentiate. On the other hand, it has been known that hair growth is promoted by treatment of a mouse with a winter protein, and that hair growth is inhibited in a mouse knock-downed with a winter gene. This means that substances promoting the movement of the hair / beta-catenin signaling present in the hair follicle stem cells can promote hair growth. Since the substances promoting hair growth through these mechanisms are not involved in the promotion of hair growth through regulation of male hormone and / or blood circulation, it is possible to prevent hair growth by inhibiting male hormones or preventing hair loss, It will be possible to improve the feminine hair loss. Gardenia extracts can stimulate hair growth by promoting stem cell wint / beta-catenin (Wnt / beta-catenin) signaling. Therefore, a preparation containing a gardenia extract promotes hair growth, and further, hair loss can be prevented and wool can be promoted. In addition, the formulation containing the gardenia extract can have hair growth promotion, hair loss prevention and wool promoting effect superior to minoxidil, which is known as a hair growth promoter, and shows almost no side effects such as sexual dysfunction or skin irritation, Can be used.

In one aspect of the invention, gardenia (梔子) is an evergreen shrub gardenia dicotyledonous happanhwa group madder neck tree Rubiaceae (Gardenia There are jasminoides . grandiflora). Gardenia fruit contains saponin and grosin, and flower has a lot of flower oil (perfume), and it is known to have fatigue recovery, hypnosis, dryness, diuretic, dressing, fever, appetite enhancement effect.

In one aspect of the present invention, a gardenia extract can be obtained by solvent extraction of a dried gardenia. The solvent includes at least one selected from the group consisting of water, an organic solvent, and a mixture of water and an organic solvent. Wherein the water comprises distilled or purified water and the organic solvent is selected from the group consisting of alcohols such as C ₁ -C ₅ lower alcohols, acetone, ether, ethyl acetate, diethyl ether, ethyl methyl ketone and chloroform But is not limited thereto.

Accelerators according to one aspect of the invention may comprise from 0.01% to 10% by weight, in particular from 0.1% to 5%, more specifically from 0.5% to 3% by weight of the gardenia extract, based on the total weight of the accelerator have. When it is included in the above range, it is not only suitable for exhibiting the intended effect of the present invention but also can satisfy both the stability and safety of the composition and may be suitably included in the above range in terms of cost effectiveness. Specifically, when the extract of the gardenia extract is less than 0.01% by weight, sufficient skin regeneration and hair growth promoting effect can not be obtained. When it exceeds 10% by weight, safety and formulation stability may be lowered.

One aspect of the present invention provides a cosmetic composition comprising a gardenia extract as an active ingredient. The cosmetic composition has a skin regenerating effect, specifically, it can heal wounds and reduce scars. In addition, the cosmetic composition promotes the activity of the hair so that the hair growth is promoted, and further, the hair loss can be prevented and the wool can be promoted.

The cosmetic composition according to the present invention may be provided in all formulations suitable for topical application. For example, it can be provided as a formulation of a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an oil phase in water, a suspension, a solid, a gel, a powder, a paste, a micro needle, a foam or an aerosol composition have. Compositions of such formulations may be prepared according to conventional methods in the art.

The cosmetic composition according to the present invention may contain, in addition to the above-mentioned substances, other ingredients which can give a synergistic effect to the main effect, so long as they do not impair the main effect. The cosmetic composition according to the present invention may further comprise a moisturizing agent, an emollient agent, a surfactant, an ultraviolet absorber, an antiseptic, a bactericide, an antioxidant, a pH adjuster, an organic or inorganic pigment, a fragrance, have. The compounding amount of the above components can be easily selected by those skilled in the art within a range not to impair the objects and effects of the present invention. The amount thereof may be 0.01 to 5% by weight, specifically 0.01 to 3% by weight, have.

One aspect of the present invention provides a pharmaceutical composition comprising a gardenia extract as an active ingredient. The pharmaceutical composition has a skin regenerating effect, and can specifically heal wounds and reduce scarring. In addition, the pharmaceutical composition promotes hair growth, promotes hair growth, further prevents hair loss, and promotes hair growth.

The pharmaceutical composition according to one aspect of the present invention may be administered orally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, and the like.

Formulations for oral administration may be in the form of tablets, pills, soft and hard capsules, granules, powders, granules, solutions, emulsions or pellets, It is not. These formulations may contain, in addition to the active ingredient, a diluent such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose or glycine, a lubricant such as silica, talc, stearic acid or polyethylene glycol, or a binder such as magnesium aluminum Silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose or polyvinylpyrrolidine). And may optionally contain pharmaceutical additives such as disintegrants, absorbents, colorants, flavors, or sweeteners. The tablets may be prepared by conventional mixing, granulating or coating methods.

Formulations for parenteral administration may be, but are not limited to, injections, drops, lozenges, ointments, gels, creams, suspensions, emulsions, suppositories, patches or spraying agents.

The effective ingredients of the pharmaceutical composition according to one aspect of the present invention will vary depending on the age, sex, weight, pathological condition and severity of the subject to be administered, route of administration, or judgment of the prescriber. Determination of the amount of application based on these factors is within the level of ordinary skill in the art and its daily dose is, for example, from 0.1 mg / kg / day to 100 mg / kg / day, more specifically from 5 mg / kg / day to 50 mg / / Day, but is not limited thereto.

One aspect of the present invention provides a food composition comprising a gardenia extract as an active ingredient. The food composition has a skin regenerating effect, and can specifically heal wounds and reduce scars. In addition, the food composition promotes the activity of the hair so that the hair growth is promoted, and further, the hair loss can be prevented and the wool can be promoted. The food composition comprises a preference food or a health food composition.

The formulation of the food composition is not particularly limited, but may be formulated into, for example, tablets, granules, powders, liquid preparations such as drinks, caramels, gels, bars and the like. The food composition of each formulation can be blended with the ingredients commonly used in the field in addition to the active ingredient without difficulty by those skilled in the art depending on the purpose of formulation or use, and synergistic effect can be obtained when the composition is applied simultaneously with other ingredients.

The dosage of the active ingredient is within the level of those skilled in the art and its daily dose is, for example, from 0.1 mg / kg / day to 5000 mg / kg / day, more specifically from 50 mg / kg / day, but it is not limited thereto, and may be varied depending on various factors such as the age, health condition, and complication of the subject to be administered.

Hereinafter, the constitution and effects of the present invention will be described in more detail with reference to examples and test examples. However, these examples and test examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited thereto.

EXAMPLES Preparation of Gardenia Extract

Dried gardenia (Gardenia 1 kg of jasminoides ) is selected and controlled according to the general rule of pharmacopoeia and the degree of the powder, and 5 to 10 times of water is added thereto and the mixture is leached at 90 to 100 ° C for 2 to 3 hours. The leached solution is concentrated under reduced pressure, then about 1/3 volume of ethanol is added to the leached solution and allowed to stand overnight in cryo. Then, according to the preparation method of the Korean Pharmacopoeia General Instructions Extract, about 330 g of the soft X-shaped periodontal extract was obtained by softening the extract. Approximately 1 g of soft-drink extract corresponds to about 3.03 g of herbal medicine.

[Experimental Example 1] Evaluation of promoting effect of stem cell /

When the Wnt signal is given to stem cells, beta-catenin migrates into the cell nucleus as a result of inhibition of the intracellular beta-catenin-inhibiting protein complex (GSK3, APC, Axin) Various genes are expressed.

Beta-catenin binds to a specific nucleotide sequence of a chromosome as a transcriptional regulator. A plasmid vector in which a Gaussia luciferase gene sequence has been artificially combined with the β-catenin adhesion sequence has been prepared, and HEK293 Lt; / RTI > cell line). The transformed cell line thus prepared is resistant to G418 antibiotic. When cultured in a medium containing G418, only the transformed cell line can be selectively cultured. The transformed HEK293 cell line having the thus obtained winter / beta-catenin signaling pathway was treated with the gardenia extract of the example, and then the expression of Gaussia luciferase was determined by the Gaussia luciferase assay kit (BioLabs) Respectively.

Specifically, the transgenic HEK293 cells, 10% bovine serum serum (Fetal bovine serum) (FBS) and G418 500μg / Dulbecco's modified Eagle's medium with ml containing; a (DMEM Gibco BRL, Gaithersburg, MD , USA) 5% CO 2 , And cultured in an incubator at 37 ° C for 24 hours. Cells cultured in 96-well plates for 24 hours were treated with 20 ppm and 50 ppm of gardenia extract, respectively, and cultured in an incubator for 48 hours. Sodium valproic acid (VPA) 16.6 ppm, 50 ppm and 83 ppm, which are widely used as inhibitors of glycogen synthase kinase 3 beta (GSK-3 beta), were used as positive control. The luciferase expression of cells cultured with the Gaussia luciferase assay kit was measured and the effect of the gardenia extract on the wint / beta-catenin signaling was evaluated. The results are shown in Fig.

As can be seen from Fig. 1, the gardenia extract promotes the wight / beta-catenin signaling of stem cells. As a result, it can be seen that the gardenia extract has an effect of stimulating skin regeneration and hair growth.

[Test Example 2] Alkaline phosphatase activity evaluation

In order to evaluate whether or not the gardenia extract can maintain the stem cell characteristics of the human dermal papilla cell line, an in vitro system was applied and the alkaline phosphatase enzyme involved in the stem cell maintenance of the cell line after application of the gardenia extract Activity.

For this evaluation, human immortalized dermal papilla (iDPc), an immortalized dermal papilla cell prepared by transducing human dermal papilla cells with vectors containing Simian virus 40 large T (SV40T) and c-myc cell) was used. The cell lines were cultured in a 5% CO ₂ incubator at 37 ° C. for 24 hours in Dulbecco's modified Eagle's medium (DMEM; Gibco BRL, Gaithersburg, MD, USA) containing 10% bovine serum serum (FBS) 100ppm of gardenia extract was treated. DMSO was used as a negative control and 83 ppm of sodium valproic acid (VPA), widely used as a glycogen synthase kinase 3 beta (GSK-3?) Inhibitor, was used as a positive control. 166 ppm was used. After 48 hours, the cells were washed twice with phosphate buffered saline (PBS), and then 300 μl of ALP assay buffer (BioVision, Catalog # K412-500) was added to the cells. Scraped and collected in an eppendorf tube. The collected cells were wakened with a probe sonicator on ice, centrifuged at 4 ° C, 12,000 rpm for 5 minutes, and the supernatant was taken. According to the manufacturer's recommended method, using the Bio-rad DC protein kit (Biorad, Cat. No. 500-0111) based on the Lowry's method, The concentration of cellular protein was measured. The alkaline phosphatase activity of the test substance was evaluated by converting the measured concentration value.

Specifically, nitrophnyl phosphate (pNPP), which is a substrate of alkaline phosphatase enzyme, is dissolved in assay buffer of pNPP tab (ALP assay kit, BioVision, Catalog # K412-500) I put it. PNPP dissolved at a concentration of 1 mM was added to each of the concentrations to 4 ~ 20 ㎕, and the standard solution was prepared by allowing the enzyme digestion reaction of pNPP to take place by adding ALP enzyme. The 96-well plate containing the test substance and the standard solution was placed in a 25 ° C incubator for 1 hour, and then the stop solution was added thereto. The absorbance at 405 nm was measured by pNPP The activity of the enzyme was determined from the amount of pNP produced by digestion. The results are shown in Fig.

As can be seen from Fig. 2, the gardenia extract increased alkaline phosphatase activity in the dermal papilla cells. In other words, it can be seen that the gardenia extract promotes hair growth by maintaining and activating the stem cell of the papilla cell line.

[Test Example 3] Evaluation of proliferation effect of dermal papilla cells

The keratin proteins that make up the hair are produced in hair follicle keratinocyte, which keratinocytes differentiate from the dermal papilla cells. Therefore, if the gardenia extract promotes the activity of the dermal papilla cells, the activity of the keratinocyte differentiated therefrom can be promoted, and furthermore, the hair growth can be promoted.

In order to evaluate the promoting effect of the gardenia extract on the dermal papilla cell activity, iDPc (human immortalized dermal papilla cell) cell line was used in the present invention. This cell line is an immortalized dermal papilla cell prepared by transduction human dermal papilla cells with vectors containing Simian virus 40 large T (SV40T) and c-myc. The cells were cultured for 24 hours in an incubator maintained at 37 ° C in 5% CO ₂ with Dulbecco's modified Eagle's medium (DMEM; Gibco BRL, Gaithersburg, MD, USA) containing 10% 20ppm and 50ppm of gardenia extract were treated. As a negative control, DMSO was used. Cell proliferation (%) was measured using WST-1 kit (Roche) after 24 hours of treatment of the test substance. The results are shown in Fig.

As shown in FIG. 3, the gardenia extract significantly increased the proliferation of the dermal papilla cells. Thus, it can be seen that the gardenia extract promotes the proliferation of dermal papilla cells and further promotes keratinocyte cell activity and hair growth.

[Test Example 4] Evaluation of hair growth promoting effect using mouse

Seven - week - old C57BL / 6 mouse females were used to determine the hair growth of the gardenia extract. After hair removal, mice and other hair were sprayed with 1% and 2% of gardenia extract for 8 weeks with 50 μl twice a day from the following day. As a negative control group, 2% minoxidil, which is a main component of the hair growth preparation, was used as a solvent and a positive control group, respectively. At the end of the test, visual evaluation based on the criteria in Table 1 and hair weight measurement after hair removal were evaluated. Figs. 4 to 6 show visual evaluation results (Fig. 4), hair weight measurement results (Fig. 5), and photographs of mice after 8 weeks of testing (Fig. 6).

As shown in the above results, hair growth promoting effect and initial hair growth promoting effect were higher in the group treated with 2% gardenia extract than in the positive control group, minoxidil. As can be seen from Fig. 4, the hair growth effect of 2% gardenia application group preceded minoxidil until the fifth test. As shown in FIG. 5 and FIG. 6, hair weight of the 2% gardenia-applied group was higher than that of the non-treated group at 8 weeks after the 6th week of the experiment, while the hairiness effect of minoxidil increased. Respectively. In other words, the gardenia extract has an excellent effect of promoting hair growth, which is preclinical, and exhibits its effect quickly.

Formulation examples of compositions according to one aspect of the present invention are described below, but may be applied to various other formulations, which are not intended to be limiting but merely illustrative of the invention.

[Formulation Example 1] Preparation of ointment

Ointment is prepared according to a conventional method with the composition shown in the following table.

ingredient Content (% by weight) Purified water Balance glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 15.0 Beta Glucan 7.0 Carbomer 0.1 Gardenia extract 5.0 Caprylic capric triglyceride 3.0 Squalene 1.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Stearyl alcohol 1.0 antiseptic Suitable amount incense Suitable amount Pigment Suitable amount beeswax 4.1

[Formulation Example 2] Production of hair lotion

A hair lotion is prepared according to a conventional method with the composition shown in the following table.

ingredient Content (% by weight) Purified water Balance glycerin 3.0 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 Gardenia extract 2.0 Caprylic capric triglyceride 3.0 Squalene 5.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 1.2 antiseptic Suitable amount incense Suitable amount Pigment Suitable amount Triethanolamine 0.1

[Formulation Example 3] Preparation of cream

Creams are prepared according to a conventional method with the composition shown in the following table.

ingredient Content (% by weight) Purified water Balance glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 45.0 Beta Glucan 7.0 Carbomer 0.1 Gardenia extract 1.5 Caprylic capric triglyceride 3.0 beeswax 4.0 Cetearyl glucoside 1.5 Sesquioleic acid sorbitan 0.9 Vaseline 3.0 antiseptic Suitable amount incense Suitable amount Pigment Suitable amount Triethanolamine 0.1

[Formulation Example 4] Tablets

The mixture of 10 mg of gardenia extract, 50 mg of soybean extract, 100 mg of glucose, 50 mg of red ginseng extract, 96 mg of starch and 4 mg of magnesium stearate is mixed with ethanol to form granules, followed by drying and tableting.

[Formulation Example 5] Health drinks

Gardenia extract ............................ 100 mg

Citric acid ................................ 1000 mg

Oligosaccharides ................................ 100 g

Taurine .................................... 1 g

Purified Water ..................................

The above components are mixed according to a conventional health drink manufacturing method, and the mixture is stirred and heated at 85 DEG C for about 1 hour, and then the solution is sterilized by filtration.

Claims (8)

delete A promoter of a mammary prefrontal uterus that activates a Wnt signal in hair follicle stem cells of a hair loss entity, which contains a gardenia extract as an active ingredient and is not a male hair loss caused by androgen.
delete 3. The method of claim 2,
The accelerator is an accelerator that prevents hair loss and promotes hair growth and wool.
3. The method of claim 2,
Wherein the gardenia extract is contained in an amount of 0.01 to 10% by weight based on the total weight of the accelerator.
delete A cosmetic composition comprising the accelerator according to claim 2.
A pharmaceutical composition for improving, preventing or treating hair loss comprising the promoter according to claim 2.

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