KR20210066419A - Composition for preventing hair loss or promoting hair growth comprising Zein and Pullulan - Google Patents

Abstract

The present invention relates to a composition for preventing hair loss or promoting hair growth comprising zein and pullulan. According to the present invention, by including zein and pullulan, which are essential active components, the differentiation of keratinocytes is promoted, thereby providing an excellent effect of preventing hair loss or promoting hair growth.

Description

Composition for preventing hair loss or promoting hair growth comprising zein and pullulan {Composition for preventing hair loss or promoting hair growth comprising Zein and Pullulan}

The present invention relates to a composition for preventing hair loss or promoting hair growth, comprising zein and pullulan, and more specifically, by including essential active ingredients zein and pullulan, the expression of a gene encoding surreisin is suppressed, It relates to a composition that promotes the expression of a gene encoding loricrin and exhibits an excellent effect of preventing hair loss or promoting hair growth.

Human hair grows from epithelial cells-derived hair follicles and hair follicles. Above the hair follicle, there are Dermal Papilla Cells that dominate the hair, and there are Hair Mother Cells directly above the hair follicle, which continue to divide and produce new hair and push it up.

Each hair of the human body operates and proceeds in an independent cycle, and as a specific cycle, it grows and falls out while going through the anagen, catagen, and telogen. In general, the hair loss phenomenon can be divided into a general telogen hair loss phenomenon, in which hair falls out in a telogen state, and a growth phase alopecia phenomenon, which is an abnormal hair loss phenomenon. In particular, the latter is called alopecia.

The cause of hair loss is not yet clearly identified, but genetic causes are largely located, and it is known that it is caused by poor blood circulation in the scalp, excessive action of male hormones, excessive sebum secretion, aging, stress, and the like. Various hair loss prevention and hair growth agents are being developed according to these various causes, specifically, vasodilators such as capronium chloride, minoxidil and extracts of various plants to circulate sufficient blood to the scalp, and the action of male hormones that cause hair loss There are female hormones such as estrogen, estraziol, and progesterone, or male hormone activation inhibitors such as pentadecanoic acid and finasteride.

However, most of the hair growth agents show some effect in preventing hair loss, but the effect related to hair growth was not very satisfactory, and Korean Patent Publication No. 10-1467568 contains minoxidil, an emollient and a skin retention agent. Although the hair growth effect of the composition is described, the prior art including the above patent does not predict whether the above effect is exerted due to any active ingredient, and there is a problem that it still does not show a satisfactory effect.

Accordingly, there is a need for the development of a composition containing essential active ingredients to exhibit an excellent hair loss prevention or hair growth promoting effect.

Republic of Korea Patent Publication No. 10-1467568 (Announcement date of 2014.12.02)

The present invention has been devised to solve the above problems, and an object of the present invention is to contain essential active ingredients zein and pullulan, thereby suppressing the expression of a gene encoding surreyosin, while encoding loricrin. To provide a composition that promotes gene expression and exhibits an excellent hair loss prevention or hair growth promoting effect.

In order to achieve the above object of the present invention, the present invention provides a composition for preventing hair loss or promoting hair growth, comprising zein and pullulan as active ingredients.

According to the present invention, by including zein and pullulan, which are essential active ingredients, the expression of the gene encoding ceryaline is suppressed, while the expression of the gene encoding loricrin is promoted, thereby providing excellent hair loss prevention or hair growth promoting effect. to provide.

1 is a view evaluating the effect on human dermal papilla cell growth according to an embodiment of the present invention.
Figure 2 is a view evaluating the effect on hair follicle length growth according to an embodiment of the present invention.
3 is a view confirming the expression level of S100A7 in keratinocytes according to an embodiment of the present invention.
4 is a view confirming the expression level of loricrin mRNA in keratinocytes according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail with reference to examples and drawings.

The present invention provides a composition for preventing hair loss or promoting hair growth comprising zein and pullulan as active ingredients.

The composition of the present invention may be a pharmaceutical composition or a cosmetic composition.

In addition, the present invention provides a use for preventing hair loss or promoting hair growth of the active ingredient contained in the composition of the present invention.

In addition, the present invention provides a method for preventing hair loss or promoting hair growth, comprising administering to a mammal, including or not including a human, in need of the active ingredient contained in the composition of the present invention.

The present invention also provides the use of zein and pullulan to produce a pharmaceutical formulation having the efficacy of preventing or treating alopecia.

In addition, the present invention provides a pharmaceutical composition for preventing or treating alopecia, preventing or promoting hair growth, comprising zein and pullulan as active ingredients.

The alopecia (or hair loss disease) may be alopecia areata, hereditary androgenetic alopecia, telogen alopecia, traumatic alopecia, alopecia on the hair, pressure alopecia, genital alopecia, alopecia pityis, syphilitic alopecia, seborrhea It may be one or more selected from alopecia, symptomatic alopecia, non-scarring alopecia, alopecia scar, and alopecia congenital.

Zein, a kind of prolamin, a protein contained in soybeans, corn, etc., is a protein extracted from corn, and is known as a material with excellent film formation and oxidation inhibitory properties. On the other hand, pullulan, a type of α-glucan, is a high molecular biopolymer obtained by fermenting starch with black yeast (Aureobasidium pullulans ). Three glucose molecules connected by α-1,4 glycosidic bonds. It may include a trisaccharide consisting of maltotriose (maltotriose). It is hydrophilic, so it is easily soluble in water, but insoluble in alcohol. It is known as a material having excellent emulsion stability and excellent film formation and adhesion.

Hair follicles are specifically, highly nerve-dense peripheral organs that are formed and developed by epithelio-mesenchymal interactions between epidermal keratinocytes specially differentiated into hair follicles and dermal cell clusters that form dermal papilla. . Therefore, in order to prevent hair loss or promote hair growth, it is necessary to promote the differentiation of keratinocytes.

On the other hand, Psoriasin (S100 calcium-binding protein A7), a product encoded by the S100A7 gene, is known as a protein that inhibits the differentiation of keratinocytes and induces an inflammatory response, and loricrin is It is a substance necessary for the differentiation of keratinocytes to form stable keratin. Therefore, in order to promote the differentiation of keratinocytes, it is necessary to promote the expression of the gene encoding loricrin while suppressing the expression of the gene encoding the ceryaline.

Accordingly, the present inventors recognized that there is no study on the blood circulation improvement effect when zein and pullulan are combined to apply to the hair, and the effect of preventing hair loss or promoting hair growth through this, recognizing that zein and pullulan are used as active ingredients A composition for preventing hair loss or promoting hair growth was completed, including. In particular, the present inventors have discovered that zein and pullulan have the effect of promoting the differentiation of keratinocytes, and specifically, zein and pullulan inhibit the expression of a gene encoding ceryaisin while encoding loricrin The present invention was completed by discovering that there is an effect of promoting the differentiation of keratinocytes by promoting the expression of the keratinocytes.

In the present invention, the zein is replaced with gliadin, hordein, secalin or kafirin, which are other types of prolamin, or gliadin, hordein ( hordein), secalin, or kafirin may be mixed and used, but the use of zein is most preferable in terms of S100A7 expression and loricrin expression in keratinocytes.

The solvent that can be used to extract zein and pullulan is an organic solvent, and one selected from methanol, n-hexane, chloroform, butanol, hexane, ethyl acetate, dimethyl formamide, tetrahydrofuran, chloroform, diethyl ether and butanol. More than one can be used, and it is preferable to use a lower alcohol having 1 to 4 carbon atoms and water.

As described above, the composition of the present invention may be a pharmaceutical composition, and the pharmaceutical composition may include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil and the like. It also includes diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and the like.

The pharmaceutical composition may be formulated in the form of oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, parenteral preparations, injection preparations, and topical administration preparations according to conventional methods.

Among oral formulations, oral solid formulations include tablets, pills, powders, granules, capsules, and the like, and these solid formulations include at least one excipient, for example, starch, calcium carbonate, sucrose. Alternatively, it may include lactose, gelatin, and the like, and a lubricant such as magnesium stearate and talc. Among oral formulations, oral liquid formulations include suspensions, solutions, emulsions, syrups, and the like, and may include water, diluents such as liquid paraffin, wetting agents, sweeteners, fragrances, preservatives, and the like. Parenteral preparations include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, and freeze-dried preparations. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and ethylol. injectable esters such as lactate; and the like.

In addition, the pharmaceutical composition may be administered to mammals, including or not including humans, by various routes, for example, by oral, intravenous, intramuscular, or subcutaneous injection.

The compositions of the present invention may be administered topically. Topical administration refers to direct application of an active ingredient or composition to the skin and/or hair, and includes a composition for external application to the skin. The topical composition of the present invention may be in the form of a solution, lotion, plaster, cream, ointment, liposome, spray, gel, foam, roller stick, or any other formulation routinely used in dermatology. have.

In the composition, use, and method of the present invention, the dosage of the active ingredient varies depending on the condition and weight of the patient, the degree of disease, the drug form, the route and period of administration, but may be appropriately selected depending on the case. The administration may be administered once or divided several times a day.

As described above, the composition of the present invention may be a cosmetic composition, and the cosmetic composition may be prepared in various forms according to a conventional cosmetic preparation method including the zein and pullulan as active ingredients. Specifically, the cosmetic composition includes a hair tonic, a hair cream, a hair lotion, a hair shampoo, a hair conditioner, a hair conditioner, a hair spray, a hair aerosol, a pomade, a powder, a gel, a hair pack, It can be prepared in the form of hair treatment, eyebrow hair growth agent, eyelash hair growth agent, eyelash nutrition agent or general ointment.

In addition, the cosmetic composition may include conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments, and fragrances commonly used in the field of cosmetic compositions, and may include a cosmetically acceptable carrier.

Cosmetically acceptable carriers include, but are not limited to, purified water, oils, waxes, fatty acids, fatty alcohols, fatty acid esters, surfactants, humectants, thickeners, viscosity stabilizers, chelating agents, buffers, preservatives, lower alcohols, and the like. it is not If necessary, moisturizers, anti-inflammatory agents, antibacterial agents, antifungal agents, vitamins, sunscreens, antibiotics, perfumes, and dyes may be included.

{Example}

Hereinafter, the present invention will be described in detail by way of Examples. However, the following examples are only examples to help the overall understanding of the present invention, and the content of the present invention is not limited to the following examples.

<Example 1> Extraction of active ingredients zein and pullulan

Zein for use in the test example was obtained by separation and extraction from corn, and pullulan was obtained by inoculating and culturing a culture solution of a black yeast ( Aureobasidium pullulans ) strain, and then separating and purifying polysaccharides from the fermented product from which the culture was removed. .

<Test Example 1> Assessment of the effect of zein and pullulan on the growth of human dermal papilla cells (Human Dermal Papilla Cells)

The effect of zein and pullulan according to the present invention on the growth of human dermal papilla cells (Human Dermal Papilla Cells) was evaluated using human dermal papilla cells (Human Dermal Papilla Cells, Department of Dermatology, Dankook University).

Specifically, in DMEM (Dulbecco's Modified Eagle Media, HyClone GE Healthcare) medium containing 10% FBS (Fetal Bovine Serum, Cambrex), 1% antibiotics, and 0.1% fungizone, human dermal papilla cells was inoculated into a 96-well plate to become 5 × 10 ³ cells/ml, and cultured for 72 hours at 37° C., 5% CO _{2 humid conditions.} After 24 hours, it was washed twice with DPBS (Dulbecco's Phosphate Buffer Saline, Sigma Aldrich, USA), and then replaced with a medium from which FBS was removed.

The culture medium was treated with 50 ppm of zein extracted in Example 1 alone, 50 ppm of pullulan alone, and a total of 50 ppm of zein and pullulan in combination. After culturing for 72 hours, CCK-8 ( Cell viability was confirmed with Cell Counting Kit-8, CK04-11, Dojindo Molecular Technologies, Inc, MA, USA). After incubation with CCK-8 for 2 hours, absorbance at 450 nm was measured using a microplate reader.

Under the same conditions as above, the untreated group (Control) was used as a negative control group, and a culture medium treated with 5% FBS instead of zein and pullulan was used as a positive control group. The test was repeated 3 times, and the results obtained are shown in FIG. 1 .

As can be seen in Figure 1, compared to the sample untreated group (Control), the treatment group treated with a total concentration of 50 ppm by using zein and pullulan in combination improved human dermal papilla cell growth by about 120% or more.

From the results of FIG. 1, compared to the group treated with zein alone or pullulan alone, when treated with zein and pullulan included as active ingredients in the composition of the present invention in combination, it exhibits superior dermal papilla cell growth promoting activity, It can be seen that there is an excellent hair loss prevention or hair growth promoting effect.

<Test Example 2> Evaluation of the effect of zein and pullulan on the growth of hair follicles (Hair Follicles)

The effect of zein and pullulan according to the present invention on the growth of hair follicles was evaluated in organ culture using hair follicles provided by the Department of Dermatology, Dankook University.

Specifically, hair follicles were isolated from human occipital scalp tissue provided by Dankook University's Department of Dermatology using tweezers under microscopic observation (hair follicle sample information: 57 years old, female).

The hair follicles were added to a medium based on Williams E culture medium, 2mM L-glutamine, 10ng/ml hydrocortisone, 1% antibiotics, and 0.1% fungizone. and cultured. After dispensing 500 μl of the medium in a 24-well plate, 6 samples (*2) were cultured in each well.

The zein extracted in Example 1 was treated alone at a concentration of 5 ppm in the culture solution, the pullulan extracted in Example 1 was treated alone at a concentration of 5 ppm in the culture solution, and the zein and pullulan extracted in Example 1 in the culture solution were treated alone. was co-treated at a concentration of 5 ppm.

For each experimental group, 20 follicular units were used, and the medium was exchanged every 2 to 3 days during culture. After cutting along the red line shown in FIG. 2 , the hair follicle growth length was measured. A photograph of hair follicle length growth on the 7th day taken using a stereomicroscope is shown in FIG. 2 as a representative image.

From the results of Figure 2, it can be seen that, compared to the untreated group (Control), when treated in combination with both zein and pullulan, which are components included as active ingredients in the composition of the present invention, excellent hair follicle length growth promoting efficacy is shown. have. In addition, compared to the zein alone or pullulan alone treatment group, it can be seen that when zein and pullulan are treated in combination, an excellent hair follicle length growth promoting effect is exhibited. Therefore, it was confirmed that zein and pullulan had an excellent effect of promoting hair follicle length growth, preventing hair loss, promoting hair growth, and further promoting hair health.

<Test Example 3> Confirmation of expression level of S100A7 in keratinocytes

1. Cell Culture

In Keratinocyte Media (KCM, ScienCell Research Laboratories, #2101) containing 10% FBS (ScienCell Research Laboratories , #0500), 0.3 × 10 keratinocytes (keratinocytes, ScienCell Research Laboratories, #2440) isolated from human scalp ^It was inoculated into a 6-well plate to become 5 cells/ml, and cultured for 24 hours at 37° C., 5% CO _{2 humid conditions.}

The culture medium was treated with 50 ppm of zein extracted in Example 1 alone, 50 ppm of pullulan alone, and 50 ppm of zein and pullulan in combination and cultured for 24 hours. At this time, the untreated group (Control) was used as a negative control group.

2. Confirmation of expression level of S100A7 using Real-time qPCR (RT-qPCR)

For the cultured cells, RNA of keratinocytes was extracted using QIAGEN's RNeasy mini kit, and then cDNA synthesis was performed using Invitrogen's SuperScript reverse transcriptase Ⅲ kit. Real-time PCR for gene comparison was performed using 7500 Fast Real-Time PCR using 2X TaqMan universal PCR mixture (10μl), 20X TaqMan expression assay mix (1μl), cDNA (50ng) and primers (S100A7: Hs00161488_m1). carried out.

3. Confirmation result of decrease in expression level of S100A7

The results of comparing the expression level of the S100A7 gene are shown in FIG. 3 . The results of the analysis were expressed by determining the expression rate of the untreated group as 1.02 and obtaining a relative value through comparison with the untreated group. As shown in Figure 3, compared to the untreated group (Control), when treated in combination with both zein and pullulan, components included as active ingredients in the composition of the present invention, significantly reduced the expression level of the S100A7 gene can be known In addition, it can be seen that when zein and pullulan are treated in combination, the expression level of the S100A7 gene is significantly reduced compared to the zein alone or pullulan alone treatment group. Therefore, zein and pullulan reduce the expression level of the S100A7 gene and inhibit the differentiation of keratinocytes, thereby promoting hair follicle length growth, preventing hair loss, promoting hair growth, and further promoting hair health. was confirmed.

<Test Example 4> Confirmation of the expression level of loricrin mRNA in keratinocytes

1. Cell Culture

Keratinocytes were cultured in the same manner as in the cell culture method in Test Example 3.

2. Confirmation of expression level of loricrin mRNA using real-time qPCR (RT-qPCR)

The same method as in Test Example 3 was performed, except that Loricrin: Hs01894962_s1 was used as a primer used for real-time PCR for gene comparison.

3. Confirmation result of decreased expression level of loricrin mRNA

The results of comparing the expression levels of loricrin mRNA are shown in FIG. 4 . The results of the analysis are shown by determining the expression rate of the untreated group as 1.00 and obtaining a relative value through comparison with the untreated group. As shown in FIG. 4 , when treated in combination with both zein and pullulan, which are components included as active ingredients in the composition of the present invention, the expression level of loricrin mRNA was significantly increased compared to the untreated group (Control) Sikkim can be seen In addition, it can be seen that when zein alone or pullulan alone treatment group, when zein and pullulan were treated in combination, the expression level of loricrin mRNA was significantly increased. Therefore, zein and pullulan increase the expression level of loricrin mRNA, differentiate keratinocytes, and form stable keratin, thereby promoting hair follicle length growth, preventing hair loss, promoting hair growth, and further improving hair health It was confirmed that there was an excellent effect of promoting.

Although the present invention has been described with reference to the above-mentioned preferred embodiments, various modifications and variations are possible without departing from the spirit and scope of the invention. Accordingly, it is intended that the appended claims cover such modifications and variations as fall within the scope of the present invention.

Claims (11)

A pharmaceutical composition for preventing hair loss or promoting hair growth, comprising zein and pullulan as active ingredients. According to claim 1,
The composition is a pharmaceutical composition for preventing or treating alopecia, preventing hair loss or promoting hair growth. 3. The method of claim 2,
The alopecia is alopecia areata, hereditary androgenetic alopecia, telogen alopecia, traumatic alopecia, alopecia on the hair, pressure alopecia, genital alopecia, alopecia pityroides, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, nonscarring alopecia, scar alopecia , And at least one selected from congenital alopecia, a pharmaceutical composition for preventing hair loss or promoting hair growth. According to claim 1,
The zein and pullulan promote the differentiation of keratinocytes, a pharmaceutical composition for preventing hair loss or promoting hair growth. 5. The method of claim 4,
The differentiation of the keratinocytes is by any one or more mechanisms of a mechanism for suppressing the expression of a gene encoding surreisin and a mechanism for promoting the expression of a gene encoding loricrin, a pharmaceutical composition for preventing hair loss or promoting hair growth . 6. The method according to any one of claims 1 to 5,
The composition is a composition for external application to the skin, a pharmaceutical composition for preventing hair loss or promoting hair growth. A cosmetic composition for preventing hair loss or promoting hair growth, comprising zein and pullulan as active ingredients. 8. The method of claim 7,
The zein and pullulan promote the differentiation of keratinocytes, a cosmetic composition for preventing hair loss or promoting hair growth. 9. The method of claim 8,
The differentiation of the keratinocytes is by any one or more mechanisms of a mechanism of suppressing the expression of a gene encoding surreisin and a mechanism of promoting the expression of a gene encoding loricrin, a cosmetic composition for preventing hair loss or promoting hair growth . 10. The method according to any one of claims 7 to 9,
The composition is a composition for external application to the skin, a cosmetic composition for preventing hair loss or promoting hair growth. 11. The method of claim 10,
The formulation of the composition is hair tonic, hair cream, hair lotion, hair shampoo, hair conditioner, hair conditioner, hair spray, hair aerosol, pomade, powder, gel, hair pack, hair treatment, eyebrow hair growth agent, eyelash hair growth agent, eyelash nutrition agent, And selected from the group consisting of a general ointment, a cosmetic composition for preventing hair loss or promoting hair growth.

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