KR102467931B1 - Brevibacillus borstelensis strain and its use for improving hair or scalp condition - Google Patents

Abstract

It relates to novel microorganisms, their lysates, culture solutions, extracts, fermented solutions, and their use for improving hair or scalp conditions. According to the novel Brevibacillus bosterensis strain according to one aspect, since it has effects of promoting hair growth, preventing hair loss, and strengthening hair roots, it can be variously applied for improvement, prevention, or treatment of hair or scalp conditions.

Description

Brevibacillus borstelensis strain and its use for improving hair or scalp condition {Brevibacillus borstelensis strain and its use for improving hair or scalp condition}

It relates to novel microorganisms, their lysates, culture solutions, extracts, fermented solutions, and their use for improving hair or scalp conditions.

In the human body, hair grows from hair follicles on the scalp, and cycles of growth (anagen), catagen (catagen), and telogen (telogen) are continuously repeated throughout life and go through generation, growth, and shedding (Paus et al. , J Invest Dermatol. 113, pp.523-532, 1999). In the catagen phase of the hair follicle, the growth of the hair follicle is stopped, and apoptosis and reorganization of the extracellular matrix occur. Therefore, as the number of hair follicle cells in the catagen and telogen phases of the hair follicle increases, the hair falls out and does not grow back. .

Hair is divided into the visible hair shaft and the invisible hair root, and among the hair roots, the root from which hair grows is called the hair bulb, and the recessed part at the bottom is called the hair papilla (Fig. One). The dermal papilla is the most important part in the development of hair, and has blood vessels and nerves to deliver nutrients and growth factors to promote hair growth. Since the number of hairs decreases due to the death and growth reduction of dermal papilla cells, proliferation of dermal papilla cells is required to maintain and increase hair.

Vascular endothelial growth factor (VEGF) is a glycoprotein that promotes angiogenesis by inducing the growth of vascular endothelial cells. VEGF induces proliferation of dermal papilla cells by attaching to VEGF receptor-2 (VEGFR-2) present in the dermal papilla and inducing the VEGF-2/ERK pathway (Wei Li et al., Exp Cell Res. 2012 Aug 15;318( 14):1633-40). Therefore, the increase in VEGF gene expression in dermal papilla cells can be of great help in inducing hair growth.

Recently, as environmental pollution and urban stress become more serious, as the number of modern people with alopecia increases, interest in materials capable of improving them is increasing. Therefore, an attempt was made to invent a substance capable of strengthening the hair root and helping to prevent hair loss by inducing hair growth by increasing the expression of growth factors in the dermal papilla cells responsible for the start of hair production in the hair root.

In this regard, Korean Patent Registration No. 10-2102578 discloses a cosmetic composition for promoting hair growth or hair growth containing a plurality of compounds. However, there are not many studies on microorganisms that can improve hair or scalp conditions.

Probiotics is a collective term for microorganisms that have a beneficial effect on the human body, and refers to microorganisms that benefit the body. Most probiotics known to date are known as lactic acid bacteria. Probiotics have been reported to produce effective effects through various beneficial effects on the human body, but studies on the interaction between scalp flora and scalp are insufficient.

Therefore, it is necessary to develop a fungus on the scalp that can be usefully used for scalp-related conditions.

One aspect is to provide a Brevibacillus borstelensis strain.

Another aspect is to provide a lysate of the strain, a culture medium, or an extract of the culture medium.

Another aspect is to provide a composition comprising an extract of a Brevibacillus borstelensis strain, a lysate thereof, a culture medium, or a culture medium.

Another aspect provides a method for preventing, ameliorating, or treating a condition in a subject comprising treating or administering to a subject in need thereof an effective amount of a composition described above.

One aspect provides a Brevibacillus borstelensis strain.

The strain may be a strain belonging to the Brevibacillus sp .

The strain comprises a 16S rRNA having at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% sequence identity with SEQ ID NO: 1 it may be The strain may contain the 16S rRNA of SEQ ID NO: 1.

As used herein, the term "sequence identity" refers to the degree of identity of amino acid residues or bases between sequences after aligning both sequences in a specific comparison region so as to match each other as much as possible. Sequence identity is a value measured by optimally aligning and comparing two sequences in a specific comparison region, and a part of the sequence in the comparison region may be added or deleted compared to a reference sequence. Percentage sequence identity can be calculated by, for example, comparing two optimally aligned sequences across the comparison region, determining the number of positions in which identical amino acids or nucleic acids occur in both sequences to obtain the number of matched positions. dividing the number of matched positions by the total number of positions within the comparison range (i.e., range size), and multiplying the result by 100 to obtain a percentage of sequence identity. The percent of sequence identity can be determined using a known sequence comparison program, examples of which include BLASTN (NCBI), CLC Main Workbench (CLC bio), MegAlign™ (DNASTAR Inc), and the like.

The Brevibacillus bostelensis ( Brevibacillus borstelensis ) The strain may be a strain deposited with accession number KCCM12815P.

The strain may have an effect of improving hair or scalp conditions. For example, the hair or scalp condition improvement may be one or more of hair growth promotion, hair loss prevention, hair root strengthening, scalp lipid inhibition, dandruff prevention, and scalp inflammation inhibition. In some embodiments, the improvement of hair or scalp conditions may be one or more of promoting hair growth, preventing hair loss, and strengthening hair roots.

The strain may be isolated. In this specification, the term “isolated” means something that is artificially separated and available that does not exist in nature. The strain was artificially isolated and deposited under the accession number. The strain may be isolated from human skin. The strain may be isolated from human scalp.

Another aspect provides a lysate of the strain, a culture medium, or an extract of the culture medium.

Details of the strain are as described above.

As used herein, the term "lysate" may refer to a product obtained by crushing the cell wall of a strain by applying chemical or physical force to the strain itself.

As used herein, the term "culture medium" may be used interchangeably with "culture supernatant", "conditioned culture medium" or "conditioned medium", and provides nutrients so that Brevibacillus bosterensis strains can grow and survive in vitro. It may mean the entire medium including the strain obtained by culturing the strain for a certain period of time in a medium that can be supplied, metabolites thereof, and extra nutrients. In addition, the culture solution may mean a culture solution obtained by removing the cells from the cell culture solution obtained by culturing the strain. On the other hand, the liquid from which the cells are removed from the culture solution is also called "supernatant", and the culture solution is left still for a certain period of time to take only the liquid of the upper layer except for the part sunk in the lower layer, remove the cells through filtration, or centrifuge the culture solution to It can be obtained by removing the precipitate and taking only the upper liquid. The "cell" refers to the strain itself of the present invention, and includes the strain itself isolated and selected from a skin sample or the like, or a strain isolated from a culture medium by culturing the strain. The cells can be obtained by centrifuging the culture solution and taking the part that has sunk to the lower layer, or it can be obtained by leaving it for a certain period of time and then removing the upper liquid because it sinks to the lower layer of the culture medium by gravity.

The culture solution may include the culture solution itself obtained by culturing the strain, its concentrate or lyophilisate, or the culture supernatant obtained by removing the strain from the culture solution, its concentrate or lyophilisate.

The culture medium may be obtained by culturing the Brevibacillus bosterensis strain in a medium (eg, R2A medium) at any temperature above 10 ° C or below 40 ° C for a certain period of time, for example, 4 to 50 hours. have.

In one embodiment, the culture supernatant of the strain may be obtained by centrifuging or filtering the strain culture medium to remove the strain.

In another embodiment, the concentrate may be obtained by concentrating the supernatant obtained after filtering the strain culture medium itself, or the culture medium using a centrifugal separation or filter.

Culture medium and culture conditions for culturing the Brevibacillus bosterensis strain can be appropriately selected or modified by those skilled in the art.

As used herein, the term "culture broth extract" refers to an extract obtained from the culture medium or a concentrate thereof, and may include an extract, a dilution or concentrate of the extract, a dried product obtained by drying the extract, or a crude or purified product thereof, or a fraction obtained by fractionating the same. can

Another aspect provides the use of a Brevibacillus bosterensis strain, a lysate, culture, or extract of the strain. Specifically, it provides a composition comprising a Brevibacillus bosterensis strain, a lysate thereof, a culture medium, or an extract of the culture medium.

In the use or composition, the Brevibacillus bosterensis strain may be a Brevibacillus bosterensis strain according to the above aspect. In one embodiment, the strain has at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% sequence identity with SEQ ID NO: 1 It may include 16S rRNA having In another embodiment, the strain may be one containing the 16S rRNA of SEQ ID NO: 1. In another embodiment, the strain may be a strain deposited with accession number KCCM12815P.

Uses of the strain may include improving hair or scalp condition.

The hair or scalp condition improvement may be one or more of hair growth promotion, hair loss prevention, hair root strengthening, scalp lipid inhibition, dandruff prevention, and scalp inflammation inhibition. In some embodiments, the improvement of hair or scalp conditions may be one or more of promoting hair growth, preventing hair loss, and strengthening hair roots.

As used herein, the term "hair growth" means that hair is generated and grown.

In this specification, the term "hair growth promotion" is used interchangeably with "hair growth promotion", and means to increase the specific gravity of anagen hair in total hair by promoting hair generation and growth. This includes not only stimulating new hair growth, but also allowing existing hair to grow healthily.

As used herein, the term "alopecia" refers to a phenomenon in which hair falls off from the scalp or a condition in which hair becomes sparse or thin. The hair loss may be caused by a decrease in the specific gravity of anagen hair. The hair loss may be caused by extrinsic factors or endogenous factors. The extrinsic factors refer to various external factors, such as ultraviolet rays, and the endogenous factors include oxidative stress due to cell death of dermal papilla cells caused by telomeres and cell function degradation, etc., which occur over time. The hair loss is alopecia areata, hereditary androgenetic alopecia, telogen effluvium, traumatic alopecia, hair loss due to trichotillomania, pressure alopecia, anagen alopecia, pityriasis It may be one or more selected from alopecia, syphilitic alopecia (alopecia syphlltiac), seborrheic alopecia (alopecia seborrhecia), symptomatic alopecia, scarring alopecia, and congenital alopecia, but is not limited thereto.

As used herein, the term "anti-hair loss" means to prevent or suppress hair loss. Therefore, the prevention of hair loss may include hair loss improvement, hair loss prevention, hair loss treatment, and the like.

In this specification, the term "hair root strengthening" refers to all actions that strengthen and strengthen the hair root part of the hair. For example, hair roots can be strengthened by proliferation of dermal papilla cells, promotion of nutrient delivery to dermal papilla cells, and increased expression of growth factors in dermal papilla cells.

As used herein, the term "scalp lipid" refers to lipids produced in the scalp. Excessive accumulation of lipids on the scalp causes problems such as shine, dandruff, stinging, and itching. When sebum is excessively secreted, a lipid film is formed on the scalp. When the lipid film is excessively formed on the scalp, the propagation of anaerobic bacteria and the production of fatty acids increase, which can cause various troubles, seborrheic scalpitis, odor, hair loss, and the like. The lipid may mean a lipid peroxide.

As used herein, the term "suppressing scalp lipids" refers to any action that reduces or inhibits the production of lipids in the scalp. Prevention of dandruff, improvement or suppression of scalp inflammation and itching due to sebum, and prevention of hair loss due to lipid peroxide can be obtained through inhibition of scalp lipids.

As used herein, the term "dandruff" refers to a phenomenon in which epidermal detachment occurs on the scalp and dead skin cells appear conspicuously. Dandruff can be accompanied by itching.

As used herein, the term "anti-dandruff" means preventing or inhibiting the production of dandruff. Therefore, the anti-dandruff may mean improving dandruff, preventing dandruff, and the like.

As used herein, the term "scalp inflammation" means inflammation occurring in the scalp. The scalp inflammation may include any one or more of seborrheic dermatitis, pruritus, dry eczema, erythema, urticaria, psoriasis, drug rash, and scalp acne.

As used herein, the term “suppressing scalp inflammation” refers to any action that reduces or improves scalp inflammation or prevents or suppresses the occurrence of scalp inflammation.

As used herein, the term "prevention" includes inhibiting the occurrence of a disease.

As used herein, the term "treatment" includes the inhibition, alleviation, or elimination of the development of a disease.

The composition according to one aspect may proliferate human follicular dermal papilla cells (HFDPC) by including a Brevibacillus bosterensis strain, a lysate thereof, a culture medium, or an extract of the culture medium. Therefore, the composition may exhibit a hair root strengthening effect, an hair loss preventing effect, and a hair growth promoting effect.

The composition according to one aspect may increase the expression of vascular endothelial growth factor (VEGF) by including a Brevibacillus bosterensis strain, a lysate thereof, a culture medium, or an extract of the culture medium. The increase in the expression of growth factors such as VEGF promotes the delivery of nutrients to dermal papilla cells and induces the growth of hair root cells, thereby preventing hair loss and promoting hair growth as well as strengthening hair roots. Therefore, the composition may exhibit a hair root strengthening effect, an hair loss preventing effect, and a hair growth promoting effect.

In another embodiment, the strain may exhibit a synergistic effect when used together with other strains having an effect of improving hair or scalp condition, for example, a strain belonging to the genus Brevibacillus. The strains belonging to the genus Brevibacillus include, for example, Brevibacillus agri, Brevibacillus antibioticus, Brevibacillus aydinogluensis, Brevibacillus brevis, Brevibacillus centrosporus, Brevibacillus choshinensis, Brevibacillus fluminis, Brevibacillus formosus, Brevibacillus fortis, Brevibacillus fulvus, Brevibacillus gelatini, Breviginsbisillus engs. halotolerans, Brevibacillus invocatus, Brevibacillus laterosporus, Brevibacillus levickii, Brevibacillus limnophilus, Brevibacillus massiliensis, Brevibacillus nitrificans, Brevibacillus panacihumi, Brevibacillus parabrevis, Brevibacillus porteri, Brevibacillus reuszeri, Brevibacillus schisleri, Brevibacillus sediminis, or Brevibacillus sediminis .

The composition is 0.001% to 80% by weight, for example, 0.01% to 60% by weight, 0.01% to 40% by weight, 0.01% to 30% by weight, 0.01% to 20% by weight, based on the total weight of the composition. %, 0.01% to 10%, 0.01% to 5%, 0.05% to 60%, 0.05% to 40%, 0.05% to 30%, 0.05% to 20%, 0.05% to 10% by weight, 0.05% to 5% by weight, 0.1% to 60% by weight, 0.1% to 40% by weight, 0.1% to 30% by weight, 0.1% to 20% by weight, 0.1% by weight % to 10% by weight, or 0.1% to 5% by weight of the strain, a lysate thereof, a culture medium, or an extract of the culture medium.

As used herein, the term "included as an active ingredient" means that the strain of the present specification, its lysate, culture medium, or extract of the culture medium is added to the extent that the above-mentioned effects can be exhibited, and drug delivery and stabilization, etc. It means that it is formulated in various forms by adding various components as subcomponents for the purpose.

The composition may be in a liquid or dry state. In one embodiment, the composition may be in the form of a dry powder.

A drying method for preparing the composition in a dry state may use a method commonly used in the art, and is not particularly limited. Non-limiting examples of the drying method include an air drying method, a natural drying method, a spray drying method, a freeze drying method, and the like. These methods may be used alone or at least two methods may be used together.

The composition may contain an additive in an effective amount sufficient to reduce deterioration of the strain, its lysate, culture medium or extract of the culture broth. The additive may be, for example, a binder, but is not limited thereto.

The composition may further include a cosmetically, pharmaceutically or food-acceptable carrier. The composition may be formulated with a carrier and provided as cosmetics, drugs, food additives, and the like.

The composition may be a cosmetic composition.

The cosmetic composition may further include ingredients commonly used in cosmetic compositions, functional additives, etc. in addition to the active ingredients disclosed herein, such as antioxidants, stabilizers, solubilizers, surfactants, dispersants, preservatives, and vitamins. , conventional adjuvants such as pigments, flavors, and the like, and carriers.

The cosmetic composition is not particularly limited to a specific formulation, and the formulation may be appropriately selected according to the purpose. The cosmetic composition may have, for example, a solubilizing formulation, an emulsifying formulation, or a dispersion formulation. The cosmetic composition may have, for example, a softening lotion, a nutrient lotion, a massage cream, a nutrient cream, an essence, a pack, a gel, an ampoule, or a skin-adhesive cosmetic formulation.

The cosmetic composition may be a formulation that can be used for hair or scalp. The cosmetic composition, for example, hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, hair treatment, hair cream, hair nutrition cream, hair moisture cream, hair massage cream, hair Wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair oil, hair drying agent, hair preservative, hair dye, hair waving agent, hair bleach, hair gel, hair glaze, hair dressing, hair lacquer, It may be in the form of a hair moisturizer, hair mousse or hair spray, specifically shampoo, cleanser, conditioner, treatment, hair pack, rinse, scalp pack, tonic, essence, lotion, cream, oil, mist, spray, or hair gel may be a formulation.

The composition may be a composition for external application for skin.

In the present specification, the external skin preparation may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal delivery patch, drug-containing bandage, lotion, or a combination thereof. The external skin preparation is a component usually used in external preparations for skin such as cosmetics or pharmaceuticals, for example, water-based components, oil-based components, powder components, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, and fragrances. , colorants, various skin nutrients, or combinations thereof and may be suitably blended as needed. The skin external preparation, metal sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid; drugs such as caffeine, tannin, bellapamil, licorice extract, glabridin, hot water extract of calin fruit, various herbal medicines, tocopherol acetate, glycylitnic acid, tranexamic acid and its derivatives or salts; Vitamin C, magnesium ascorbyl phosphate, ascorbic acid glucoside, arbutin, kojic acid; Sugars, such as glucose, fructose, and trehalose, etc. can also be mix|blended suitably.

The composition may be a pharmaceutical composition.

The pharmaceutical composition may additionally include a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, mannitol, or a combination thereof. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be calcium carboxymethylcellulose, sodium starch glycolate, calcium monohydrogen phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

The pharmaceutical composition may be formulated for oral or parenteral administration. Oral dosage forms may be granules, powders, liquids, tablets, capsules, dry syrups, and the like. Parenteral dosage forms may be injections, ointments, and the like.

The composition may be a health functional food composition.

The health functional food composition may be used alone, or in combination with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active ingredient can be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment). In general, when preparing food or beverage, the composition of the present specification may be added in an amount of 50 parts by weight or less, or 15 parts by weight or less based on the raw material. There is no particular limitation on the type of health functional food. Among the types of health functional foods, beverage compositions may contain various flavoring agents or natural carbohydrates as additional components, like conventional beverages. The natural carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; and polysaccharides such as dextrins and cyclodextrins; It is a sugar alcohol, such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The health food composition may also contain nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, and carbonated beverages. carbonation agent used, or a combination thereof. The health functional food composition may also contain natural fruit juice, fruit juice beverages, fruit flesh for preparing vegetable beverages, or a combination thereof.

Another aspect provides a method for preventing, ameliorating, or treating a condition in a subject comprising treating or administering to a subject in need thereof an effective amount of a composition described above.

The condition of the subject may be a hair or scalp condition, and may specifically be a condition related to hair loss or a condition related to inflammation of the scalp.

The terms "administering," "introducing," and "implanting" are used interchangeably herein, and according to one embodiment, a subject by a method or route that results in at least partial localization of a composition to a desired site. It can mean the placement of a composition according to one embodiment into.

Administration may be administered by a method known in the art. Administration may be administered directly to a subject by any means, for example, by routes such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. can The administration may be administered systemically or locally.

The subject may be a mammal, such as a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be a subject in need of hair or scalp condition improvement, for example, a subject in need of hair growth promotion, hair loss prevention, hair root strengthening, scalp lipid inhibition, dandruff prevention, or scalp inflammation inhibitory effect.

The administration is 0.1 mg to 1,000 mg per day of the composition according to one embodiment, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg , 5 mg to 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg may be administered. However, the dosage may be prescribed in various ways depending on factors such as formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and reaction sensitivity, and those skilled in the art can Dosage can be appropriately adjusted in consideration of these factors. The number of administrations can be once a day or two or more times within the range of clinically acceptable side effects, and administration can be performed at one or two or more sites, daily or at intervals of 2 to 5 days. The number of administration days may be administered from 1 day to 30 days per treatment. If necessary, the same treatment can be repeated after a titration period. For non-human animals, the same dosage per kg as for humans is used, or the above dosage is converted by the volume ratio (eg, average value) of the organ (heart, etc.) between the target animal and the human. A single dose can be administered.

According to the novel Brevibacillus bosterensis strain according to one aspect, since it has effects of promoting hair growth, preventing hair loss, and strengthening hair roots, it can be variously applied for improvement, prevention, or treatment of hair or scalp conditions.

1 is a diagram showing the structure of hair and dermal papilla cells.
2 is a result confirming the efficacy of increasing the proliferation capacity of dermal papilla cells.
3 is a graph showing the degree of VEGF gene expression in dermal papilla cells.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.

Example 1. Isolation and identification of strains

A sample was obtained by washing the scalp of a male in his twenties with a healthy scalp with sterile distilled water. The samples were inoculated into R2A (Reasoner's 2A) liquid medium or solid medium (manufacturer: Difco). After culturing in a 35° C. incubator for 48 hours after inoculation, 100 formed colonies were cultured in pure isolation and re-cultured in a 35° C. incubator for 48 hours. 16S rRNA sequence identification was performed on colonies completed with culture. The primers used at this time were designed to amplify in response only to bacteria. PCR amplification was carried out in 30 cycles of 95 ° C for 1 minute, 55 ° C for 1 minute, and 75 ° C for 1 minute and 30 seconds, and finally maintained at 72 ° C for 8 minutes and then stored at 4 ° C. After the PCR reaction, the DNA sequences of the isolated and cultured species were determined using ABI-3730XL (ABI, USA).

When the nucleotide sequence of the 16S rRNA region determined in the isolated and cultured microbial colony was compared and analyzed with other strains registered through the BLAST program provided on the homepage of the National Center for Biotechnology Information (NCBI), homology 97 Only novel species of less than % were selected, and among them, a new microorganism Brevibacillus borstelensis strain (hereinafter referred to as "SP-1") having 98% or less homology was selected. The selected SP-1 strain was deposited with the Korea Microorganism Conservation Center on October 28, 2020, and was given accession number KCCM12815P. SP-1 strain has a 16S rRNA sequence of SEQ ID NO: 1 (complementary DNA).

Experimental Example 1. Evaluating the efficacy of increasing the proliferative capacity of human dermal papilla cells (HFDPC)

In order to evaluate whether the culture of the SP-1 strain isolated in Example 1 significantly induces the proliferation of dermal papilla cells, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyl-2H-tetrazolium bromide) assay was performed.

First, human dermal papilla cells (HFDPC) were dispensed in 2x10 ⁴ cells/well in a 96-well plate, and then cultured for 24 hours in an incubator at 37°C and 5% CO ₂ conditions. Then, after replacing with a medium containing nothing, 1% (w/w) of the culture medium of the SP-1 strain was added and further cultured for 24 hours. A group containing 10 ppm of minoxidil (Minoxidil, MXD, 6-piperidin-1-ylpyrimidine-2,4-diamine 3-oxide), known as a hair growth promoter, was set as a positive control group instead of the strain culture medium, and a medium containing no strain culture medium The addition group (medium con) was set as a negative control group, and the culture medium addition group (None) was also set as a control group to confirm the efficacy of the strain culture medium. After 24 hours, the medium of each well was removed, washed once using DPBS (Dulbecco's phosphate buffered saline), and then 500μl of 0.5mg/mL MTT (Sigma-aldrich, St. Louis, USA) was added to each well. medium and reacted at 37°C for 4 hours. Then, absorbance was measured at 570 nm using Victor 3 (Perkin-Elmer). The cell viability was expressed as a percentage of the measured average absorbance value, and through this, the proliferative ability of human dermal papilla cells (HFDPC) can be confirmed.

2 is a result confirming the efficacy of increasing the proliferation capacity of dermal papilla cells.

As shown in Figure 2, it was confirmed that the strain of Example 1 exhibits an excellent dermal papilla cell proliferation effect.

Therefore, it was confirmed that the SP-1 strain can be used for strengthening hair roots, preventing hair loss, and promoting hair growth.

Experimental Example 2. Evaluating the efficacy of increasing the expression of growth factor (VEGF) in human dermal papilla cells (HFDPC)

In order to evaluate whether the culture solution of the SP-1 strain isolated in Example 1 can increase the expression of VEGF, a growth factor that induces hair growth and maintains scalp health, real-time polymerase chain reaction (Quantitative Real-Time Polymerase Chain Reaction: qRT-PCR) was performed.

Specifically, after dispensing 3x10 ⁵ cells/well of human dermal papilla cells (HFDPC) in a 6-well plate, they were cultured for 24 hours under cell culture conditions. After 24 hours, the medium was discarded, washed with PBS (Phosphate buffered saline), and then the cells were starved using a medium containing FBS (Fetal bovine serum). The next day, the medium was replaced with a culture medium containing 1% (w/w) of the SP-1 strain, followed by additional culture for 24 hours. A group to which 10 ppm of minoxidil (MXD) was added was set as a positive control group, a medium addition group (media con) not containing the strain culture medium was set as a negative control group, and a culture medium addition group (None) was set as a control group. Afterwards, RNA was isolated from the cells using the RNeasy Mini Kit (Qiagen), RNA was quantified at 260 nm using a nanodrop, and then 2 μg of RNA was used for each amplifier (C1000 Thermal Cycler, Bio -Rad, USA), cDNA was synthesized. Real-time polymerase chain reaction was performed in a real-time PCR machine using a mixture in which VEGF primers and cyanine dye, Cybergreen (SYBR Green supermis, Applied Biosystems, USA) were added to the synthesized cDNA, and finally the level of VEGF gene expression evaluated. The sequence and reaction conditions of the primers used are shown in Table 1 below, and the expression level of the gene was finally analyzed through correction for the β-actin gene.

Primer name direction order sequence number reaction conditions VEGF F 5′-GTGCCCACTGAGGAGTTCAAC-3′ 2 After polymerase activation at 94°C for 5 minutes, polymerization reaction at 40 cycles at 95°C for 30 seconds, 60°C for 30 seconds, and 72°C for 30 seconds R 5′-CCCTATGTGCTGGCCTTGAT-3′ 3 β-actin F 5′-GGCCATCTCTTGCTCGAAGT-3′ 4 R 5′-GAGACCTTCAACACCCCAGC-3′ 5

Figure 3 is a graph showing the level of VEGF gene expression in dermal papilla cells. As shown in Figure 3, it was confirmed that the strain of Example 1 exhibits an excellent expression increase effect of vascular endothelial growth factor (VEGF).

Therefore, it was confirmed that the SP-1 strain can be used for strengthening hair roots, preventing hair loss, promoting hair growth, and promoting scalp health.

The above description is merely an example of the technical idea of the present invention, and various modifications and variations can be made to those skilled in the art without departing from the essential characteristics of the present invention.

Korea Microbial Conservation Center (Overseas) KCCM12815P 20201028

SEQUENCE LISTING <110> Cosmax, INC. <120> Brevibacillus borstelensis strain and its use for improving hair or scalp condition <130> PN136745 <160> 5 <170> PatentIn version 3.2 <210> 1 <211> 1420 <212> DNA <213> Brevibacillus borstelensis <400> 1 tttgatgcgg tgctatcatg cagtcgagcg agtcccttcg ggggctagcg gcggacgggt 60 gagtaacacg taggcaacct gcccgtaagc tcgggataac atggggaaac tcatgctaat 120 accggatagg gtcttctctc gcatgagagg agacggaaag gtggcgcaag ctaccactta 180 cggatgggcc tgcggcgcat tagctagttg gtggggtaac ggcctaccaa ggcgacgatg 240 cgtagccgac ctgagagggt gaccggccac actgggactg agacacggcc cagactccta 300 cgggaggcag cagtaggggaa ttttccacaa tggacgaaag tctgatggag caacgccgcg 360 tgaacgatga aggtcttcgg attgtaaagt tctgttgtca gagacgaaca agtaccgttc 420 gaacagggcg gtaccttgac ggtacctgac gagaaagcca cggctaacta cgtgccagca 480 gccgcggtaa tacgtaggtg gcaagcgttg tccggaatta ttgggcgtaa agcgcgcgca 540 ggcggctatg taagtctggt gttaaagccc ggggctcaac cccggttcgc atcggaaact 600 gtgtagcttg agtgcagaag aggaaagcgg tattccacgt gtagcggtga aatgcgtaga 660 gatgtggagg aacaccagtg gcgaaggcgg ctttctggtc tgtaactgac gctgaggcgc 720 gaaagcgtgg ggagcaaaca ggattagata ccctggtagt ccacgccgta aacgatgagt 780 gctaggtgtt gggggtttca ataccctcag tgccgcagct aacgcaataa gcactccgcc 840 tggggagtac gctcgcaaga gtgaaactca aaggaattga cgggggcccg cacaagcggt 900 ggagcatgtg gtttaattcg aagcaacgcg aagaacctta ccaggtcttg acatcccgct 960 gaccgtccta gagatagggc ttcccttcgg ggcagcggtg acaggtggtg catggttgtc 1020 gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac gagcgcaacc ctttcttta 1080 gttgccagca ttcagttggg cactctagag agactgccgt cgacaagacg gaggaaggcg 1140 gggatgacgt caaatcatca tgccccttat gacctgggct acacacgtgc tacaatggct 1200 ggtacaacgg gaagctagct cgcgagagta tgccaatctc ttaaaaccag tctcagttcg 1260 gattgcaggc tgcaactcgc ctgcatgaag tcggaatcgc tagtaatcgc ggatcagcat 1320 gccgcggtga atacgttccc gggccttgta cacaccgccc gtcacaccac gggagtttgc 1380 aacacccgaa gtcggtgagg taaccgcaag gagccagccg 1420 <210> 2 <211> 21 <212> DNA <213> artificial <220> <223> VEGF forward primer <400> 2 gtgcccactg aggagttcaa c 21 <210> 3 <211> 20 <212> DNA <213> artificial <220> <223> VEGF reverse primer <400> 3 ccctatgtgc tggccttgat 20 <210> 4 <211> 20 <212> DNA <213> artificial <220> <223> beta-actin forward primer <400> 4 ggccatctct tgctcgaagt 20 <210> 5 <211> 20 <212> DNA <213> artificial <220> <223> beta-actin reverse primer <400> 5 gagaccttca acaccccagc 20

Claims (11)

Contains the 16S rRNA of SEQ ID NO: 1 and deposited under accession number KCCM12815P, Brevibacillus belonging to the genus Brevibacillus ( Brevibacillus sp .) Brevibacillus bosterensis ( Brevibacillus borstelensis ) SP-1 strain. The strain according to claim 1, wherein the strain is isolated from the skin. A culture solution of the strain of claim 1. A cosmetic composition comprising the strain of claim 1, a lysate thereof, a culture medium, or an extract of the culture medium. The cosmetic composition according to claim 4, which is for improving hair or scalp conditions, and is one or more of hair growth promotion, hair loss prevention, and hair root strengthening. . The method according to claim 5, wherein the hair loss is alopecia areata, hereditary androgenetic alopecia, telogen effluvium, traumatic alopecia, hair loss due to trichotillomania, pressure alopecia, A cosmetic composition that is at least one selected from anagen alopecia, pityroid alopecia, syphilitic alopecia (alopecia syphlltiac), seborrheic alopecia (alopecia seborrhecia), symptomatic alopecia, scarring alopecia, and congenital alopecia. The cosmetic composition according to claim 4, which proliferates human hair papilla cells. The cosmetic composition according to claim 4, which increases the expression of vascular endothelial growth factor (VEGF). A cosmetic composition for improving hair or scalp condition comprising an extract of Brevibacillus borstelensis SP-1 strain, its lysate, culture medium, or culture medium deposited under accession number KCCM12815P, promoting hair growth, preventing hair loss, And a cosmetic composition that is at least one of strengthening hair roots. A pharmaceutical composition for promoting hair growth or preventing hair loss, comprising an extract of Brevibacillus borstelensis SP-1 strain, a lysate thereof, a culture medium, or a culture medium deposited under Accession No. KCCM12815P as an active ingredient. Brevibacillus bostelensis ( Brevibacillus borstelensis ) SP-1 strain deposited under accession number KCCM12815P, a health functional food composition for improving hair or scalp conditions, comprising a lysate, a culture medium, or an extract of the culture medium, promoting hair growth and hair loss Prevention, and at least one of hair root strengthening health functional food composition.

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