KR101919161B1 - Composition for improving condition of hair and preventing hair loss - Google Patents

Abstract

More particularly, the present invention relates to a composition for improving scalp and preventing hair loss comprising an extract of Chaetoceros gracilis, a marine microalgae, as an active ingredient, . The microalgae of the present invention, which is a microalgae for preventing hair loss and improving the scalp, is a microalgae mainly used for oyster and sea cucumber cultivation, and is used as a food for marine life. In the present invention, the microalgae have excellent antioxidative activity, anti-inflammatory activity and 5 alpha-reductase inhibitory activity, and thus it is confirmed that hair alopecia prevention and hair growth promoting effect are exhibited. Thus, Chaotoceros gracilis ) can be used as a hair loss prevention and hair growth promoting agent.

Description

[0001] The present invention relates to a composition for improving scalp and preventing hair loss,

The present invention relates to a novel composition for scalp improvement and hair loss prevention.

In recent years, hair loss symptoms are one of the biggest problems caused by increased interest in external images. Hair loss is generally regarded as a distress for men in their 40s and 50s, but hair loss patients in their 20s and 30s are soaring, and female hair loss is also on the rise. These symptoms of hair loss are caused by many environmental factors such as unbalanced eating habits, harmful air, polluted drinking water, acid rain, and stress. Accordingly, a lot of studies are under way to prevent and treat the hair loss phenomenon.

Hair loss refers to a state in which hair normally does not exist in a region where hair should be present, or a hair that naturally stops growing. Hair usually falls between 50 and 100 or more per day, and hair loss is defined as hair loss (Lee, 2005). The causes of hair loss are very diverse, and genetic causes and male hormones, such as androgens, are considered to be important factors in the development of baldness. Some cases of female hair loss are presumed to occur in the same way as male hair loss, and alopecia areata is thought to be an autoimmune disease. In addition, resting alopecia are caused by severe physical and mental stresses such as endocrine diseases, malnutrition, drug use, birth, fever, and surgery. This is because part of the hair can not be fully grown due to temporary hair loss, Hair loss occurs.

Various attempts have been made to prevent and treat such hair loss symptoms. There have been various surgical methods such as scalp flap surgery, reduction of hair loss area, tissue expanding and self-hair grafting, and hair loss prevention and hair growth promoting agent using natural herbal extract . Although the above-described surgical method can exhibit excellent effects in a short period of time, there is a great problem that the patient's economic and psychological burden on the surgery is great. Therefore, in addition to surgical methods, various external medicines are used to prevent and prevent hair loss. Usually, side effects may occur when using synthetic medicines. These side effects include loss of libido, impaired sexual function such as erectile dysfunction, and women are at risk of birth defects.

In recent years, there have been increasing studies on the use of oriental herbal extracts for preventing hair loss and promoting hair growth using natural products having no side effects. In the past, natural products used as a hair loss prevention or hair growth promoting agent are extracts of various natural products such as phthalocyanine, yellowish white, gold, hwangjeong, hwanggi, white paper, yam chrysanthemum, licorice and ginger. In particular, when used as a hair loss preventing and hair growth promoting agent by using natural products, there is no side effect that appears when used in a synthetic preparation, an unpleasant odor is not generated, and an antioxidative effect and a natural preservative effect are provided.

However, there is a disadvantage in that hair loss prevention and scalp remedy using natural products are expensive because raw materials are expensive and cost is high in extraction process, which is not economical.

Therefore, it is a microalgae mainly used for oyster and sea cucumber cultivation which is relatively easy to supply and supply large quantities of raw materials to replace hair extract prevention and scalp improvement extracts obtained from natural products, It is necessary to develop a formulation that replaces hair loss prevention and scalp improvement agents derived from synthetic materials or expensive natural materials that have been used in the past by using marine microalgae cultured in large quantities in enterprises and the like.

Korean Patent No. 10-0950437

Accordingly, the present inventors have found that Chaetoceros gracilis, which is a microalga that is mainly used as a food for marine life, is an active microleak used for oyster and sea cucumber cultivation to prevent hair loss and promote hair growth Thereby completing the present invention.

Accordingly, an object of the present invention is to provide a composition for improving scalp and preventing alopecia containing an extract of Chaetoceros gracilis, which is a microalgae, as an active ingredient.

Another object of the present invention is to provide a health functional food for improving the scalp and preventing hair loss, which comprises an extract of Chaetoceros gracilis, which is a microalgae, as an active ingredient.

Another object of the present invention is to provide a cosmetic composition containing an extract of Chaetoceros gracilis, which is a microalgae, as an active ingredient to improve scalp and prevent hair loss.

In order to achieve the above object, the present invention provides a composition for improving scalp and preventing alopecia, which comprises an extract of Chaetoceros gracilis, which is a microalgae, as an active ingredient.

In one embodiment of the present invention, the extract may be an extract obtained by using water or an organic solvent.

In one embodiment of the present invention, the microalga Chaetoceros The extract of gracilis may contain the microalgae, the culture of the microalgae, the concentrate of the culture, or the dried product of the culture.

In one embodiment of the present invention, the extract may have antioxidant activity, anti-inflammatory activity, and 5 alpha-reductase enzyme activity inhibitory activity.

In one embodiment of the present invention, the composition is selected from the group consisting of cream, gel, patch, spray, ointment, warning agent, lotion, liniment, pasta, and cataplasma as external skin preparations for pharmaceutical use Lt; / RTI >

In addition, the present invention provides a health functional food for improving the scalp and preventing hair loss, which contains an extract of Chaetoceros gracilis, which is a microalgae, as an active ingredient.

In addition, the present invention provides a cosmetic composition for improving scalp and preventing hair loss, which comprises an extract of Chaetoceros gracilis, which is a microalgae, as an active ingredient.

In one embodiment of the present invention, the cosmetic composition may be any one selected from the group consisting of hair tonic, hair nourishing lotion, scarf treatment, hair treatment, hair rinse, hair shampoo and hair lotion.

The present invention relates to a marine microalga Chaetoceros gracilis extracts were applied to prevent hair loss and to improve the scalp.

In addition, the extract of Chaetoceros gracilis , a marine microalgae that has functions to prevent hair loss and improve scalp, is a microalgae mainly used in oyster and sea cucumber cultivation. Since it is cultured in a large amount in laboratories, schools, .

In addition, the marine microalga Chaetoceros gracilis extract can contribute to cost reduction because it can be obtained at a lower cost than natural products showing hair loss prevention effect and scalp improvement effect.

Figure 1 of the present invention Chaetoceros Gracilis extracts were tested for cytotoxicity against RAW 264.7 macrophages.
Figure 2 is a graphical representation of Chaetoceros gracilis extracts from RAW 264.7 macrophages.
Figure 3 is a graphical representation of Chaetoceros gracilis extract of LPS-induced RAW 264.7 macrophages.
Figure 4 is a graphical representation of the < RTI ID = Gracilis extracts were tested for their ability to inhibit 5 alpha reductase activity.

Hereinafter, the present invention will be described in detail.

All technical terms used in the present invention are used in the sense that they are generally understood by those of ordinary skill in the relevant field of the present invention unless otherwise defined. Also, preferred methods or samples are described in this specification, but similar or equivalent ones are also included in the scope of the present invention. The contents of all publications referred to herein are incorporated herein by reference.

The present invention relates to a novel material for improving the scalp, preventing hair loss, and promoting hair growth, which comprises a marine microalgae, Chaetoceros gracilis ) extracts.

More specifically, the present invention provides a composition for improving scalp and preventing alopecia, comprising an extract of Chaetoceros gracilis, which is a microalgae, as an active ingredient.

The active ingredient, Chaetoceros gracilis, which is contained in the composition for preventing scalp improvement and hair loss provided by the present invention, is a microalgae belonging to diatoms, and is mainly used as food for marine life.

The cell size of the microalgae is characterized by a major axis of 5.8 ± 0.5 and a minor axis of 4.4 ± 0.5.

The inventors of the present invention, as a new scalp improving and anti-hair restorer, have been studying to find new materials derived from natural materials rather than conventional synthetic compounds. Chaetoceros gracilis ) microalgae have the effect of preventing hair loss and promoting hair growth.

That is, according to one embodiment of the present invention, the antioxidative and anti-inflammatory activities of the ethanol extract of Chaetoceros gracilis microalgae were analyzed, and as a result, the antioxidative activity almost similar to that of ascorbic acid used as a conventional antioxidant And it was found to have anti-inflammatory activity by effectively inhibiting the production of nitric oxide and prostaglandin E2, inflammation inducers.

According to another embodiment of the present invention, the activity of the 5 alpha (alpha) -reduction agent causing hair loss is measured by using Chaetoceros gracilis ) were inhibited by the ethanol extract of microalgae.

On the other hand, hair loss can be caused by hair loss due to disease, hair loss due to hormonal cause, hair loss due to nutritional imbalance, or hair loss due to stress. Hair loss due to the disease is caused by genetic, autoimmune disease, burn, chemotherapy, It may be caused by psoriasis, fungal infection, radiation exposure, or tumor, and hair loss due to hormonal cause may be caused by drug, before and after pregnancy, after menopause, menopause, diabetes, pituitary dysfunction, parathyroid or thyroid dysfunction.

In addition, 5α-reductase is an important enzyme that converts testosterone into 5α dihydroxy testosterone (DHT) and accelerates signaling to AR. Finasteride, an inhibitor that inhibits 5α-reductase activity, is a therapeutic agent for male pattern baldness (Sangmyung; Propecia) and benign prostatic hyperplasia (BPH).

Therefore, it is possible to inhibit the activity of 5 alpha (-) - reductase, which causes hair loss, to inhibit the inflammatory reaction, and to prevent hair loss and promote hair growth in the case of a substance having antioxidative activity.

Therefore, the microalgae of the present invention having all of these activities can be utilized as a new material for preventing hair loss and promoting hair growth.

The composition for promoting hair loss prevention and hair growth provided by the present invention comprises Chaetoceros gracilis microalgae itself, a culture of the microalgae, a concentrate of the culture or a dried product of the culture as an active ingredient can do.

The Chaetoceros gracilis extract is prepared by culturing the microalgae using a culture medium, collecting the cultured chitosan gracilis strain by centrifugation or the like, The extract may be water, including purified water, or a solvent selected from a lower alcohol having 1 to 4 carbon atoms such as methanol, ethanol, butanol, or a mixed solvent thereof. Preferably, The extract can be obtained using a mixed solvent of water and ethanol, more preferably 50 to 100% ethanol, more preferably 70% ethanol.

The composition comprising the extract of Chaetoceros gracilis of the present invention as an active ingredient can be used as a pharmaceutical composition and a cosmetic composition for preventing hair loss, improving scalp and promoting hair growth.

The cosmetic composition according to the present invention may contain not only Chaetoceros gracilis extract as an active ingredient but also components commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, pigments And customary adjuvants such as fragrances, and carriers.

In addition, the composition of the present invention may be prepared by mixing a hair or hair restorer, which has been conventionally used, in addition to the above-mentioned Chaetoceros gracilis extract, so long as it does not impair hair growth (hair loss) .

In addition, the cosmetic composition of the present invention may be manufactured in any form conventionally produced in the art, and examples thereof include hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, Hair cosmetic cream, hair cream, hair cream, hair cream, hair cream, hair cream, hair cream, hair cosmetic cream, hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, It can be manufactured in various forms such as hair wave for hair, hair decolorant, hair gel, hair glaze, hair dresser, hair lacquer, hair moisturizer, hair mousse and hair spray. Especially, hair tonic, hair nutrition lotion, scalp treatment, hair A hair conditioner, a hair conditioner, a hair conditioner, a hair shampoo, and a hair lotion. However, But is not limited thereto.

According to a preferred embodiment of the present invention, the content of the chitosan gracilis extract of the present invention is 0.00001-30 wt%, preferably 0.5-20 wt%, more preferably 1.0-10 wt% Weight%. If the content of the extract is less than 0.00001% by weight, the effect of preventing the hair loss and improving the hair growth by the extract is greatly reduced. If the content is more than 30% by weight, skin irritation may be caused.

In addition, the composition of the present invention may be a pharmaceutical composition containing chitosercosyl saccharose extract as an active ingredient.

The pharmaceutical composition of the present invention can be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A lubricant or a flavoring agent can be used.

The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile solutions suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

In one embodiment of the present invention, the extract of the present invention may be contained in an amount of 0.00001 to 30% by weight based on the total weight of the composition.

The dermatological pharmaceutical composition of the present invention is an external preparation for skin having an effect of preventing hair loss, promoting hair growth, and improving scalp, and can be used as an external preparation for skin, such as cream, gel, patch, spray, ointment, warning agent, lotion agent, liniment agent, pasta agent or cataplasma agent But the present invention is not limited thereto.

The composition of the present invention may also be a food composition. In addition to containing the active ingredient chitoserosgracillus extract, such a food composition may contain various flavors or natural carbohydrates as an additional ingredient can do.

Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. The above-described flavors can be advantageously used as natural flavorings (tau martin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.).

The food composition of the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolates, foods, confectionery, pizza, ram noodles, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes, .

In addition, the food composition may further contain various additives such as various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring agents and aggravating agents (cheese, chocolate, etc.) Organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.

The chitoserosugracylus extract, which is an active ingredient of the present invention, is a natural substance and has almost no toxicity and side effects. Therefore, it can be safely used for long-term use for the purpose of preventing hair loss, promoting hair growth and improving scalp.

The present invention also provides a health functional food for preventing hair loss, improving scalp and preventing alopecia, comprising chitoserosugracaris extract as an active ingredient.

The health functional food of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, circles, etc. for the purpose of preventing hair loss, promoting hair growth and improving scalp.

In the present invention, the term " health functional food " refers to foods manufactured and processed using raw materials or ingredients having useful functions in accordance with Law No. 6727 on Health Functional Foods. Or to obtain a beneficial effect in health use such as physiological action.

The health functional foods of the present invention may contain conventional food additives and, unless otherwise specified, whether or not they are suitable as food additives are classified according to the General Rules for Food Additives approved by the Food and Drug Administration, Standards and standards.

Examples of the items listed in the above-mentioned "food additives" include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as persimmon extract, licorice extract, crystalline cellulose, high color pigment and guar gum; L-glutamic acid sodium preparations, noodle-added alkalis, preservative preparations, tar coloring preparations and the like.

For example, a health functional food in the form of tablets may be prepared by granulating a mixture obtained by mixing the above extract as an active ingredient of the present invention with an excipient, a binder, a disintegrant and other additives by a conventional method, Alternatively, the mixture can be directly compression molded. In addition, the health functional food of the tablet form may contain a mating agent or the like if necessary.

The hard capsule of the capsule type health functional food can be prepared by filling a normal hard capsule with a mixture of the above extract of the present invention and an additive such as an excipient and the soft capsule is chitoserosgracillus extract With an excipient such as an excipient or the like, into a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.

The ring-shaped health functional food can be prepared by molding a mixture of the above extract, which is an effective ingredient of the present invention, with an excipient, a binder, a disintegrant, etc., according to a known method and, if necessary, Or the surface may be coated with a material such as starch, talc.

The granular health functional food may be prepared by granulating a mixture of the above extract of the present invention with an excipient, a binder, a disintegrant, and the like in a granular form by a known method. If necessary, a flavoring agent, ≪ / RTI >

The health functional food may be a beverage, a meat, a chocolate, a food, a confectionery, a pizza, a ramen, a noodle, a gum, a candy, an ice cream, an alcoholic beverage, a vitamin complex and a health supplement food.

The present invention also provides a method of preventing or treating hair loss, comprising administering to the mammal an extract of chitosercrosarcylic acid.

The term "mammal " as used herein refers to a mammal that is the subject of treatment, observation or experimentation, preferably a human.

The term "therapeutically effective amount " as used herein refers to the amount of active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, physician or other clinician, The amount that induces the relief of the symptoms of the disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dose and the number of administrations of the active ingredient of the present invention will vary depending on the desired effect. The optimal dosage to be administered can therefore be readily determined by those skilled in the art and will depend upon the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, , Sex and diet, time of administration, route of administration and rate of administration of the composition, duration of treatment, concurrent administration of the drug, and the like. In the treatment method of the present invention, in the case of an adult, it is preferable to administer the chitoserosugarcillis extract of the present invention at a dose of 1 mg / kg to 250 mg / kg once or several times a day.

In the treatment method of the present invention, the composition comprising the chitosanthograsillary extract of the present invention as an active ingredient can be administered orally, rectally, intravenously, intramuscularly, intraperitoneally, intramuscularly, intrasternally, transdermally, Can be administered in a conventional manner via the intradermal route.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.

< Example  1>

Marine microalgae Chitoseros Gracylis ( Chaetoceros gracilis )  Recovery and extraction

Marine microalgae of Quito three Los Gras-less chamber (Chaetoceros gracilis) extract room Quito three Los gras from the strain deposited with the engine for the production of a less (Chaetoceros gracilis) the obtained centrifuged 15 minutes centrifuged at 3,000rpm to precipitate the fine separator by The algae chitosan gracilis was recovered, and 70% ethanol solution was added thereto in an amount (1000 g; 1 L) corresponding to 5 times the weight of the algae (200 g) , And the extract was filtered and concentrated to be used as a sample in the following experimental example.

< Experimental Example  1>

Chitoseros Gracilis ( Chaetoceros gracilis) of  Antioxidant activity measurement for scalp improvement, prevention of hair loss, and therapeutic effect

Radical formation leading to diseases of scalp and hair is known to cause hair loss. Therefore, when the activity of the enzyme that inhibits radical formation and induces oxidation is inhibited, hair loss can be prevented and treated.

Thus, the present inventors have found that Chaetoceros The degree of DPPH free radical scavenging activity was measured in order to confirm the antioxidative activity of gracilis extract. That is, the sample of the present invention (microalga Chaetoceros gracilis 500 μL of ethanol and 250 μL of 0.5 mM 1,1-diphenyl-2-picrylhydrazyl (DPPH) / ethanol solution were added, and the mixture was allowed to stand at room temperature for 30 minutes at 25 ° C., The absorbance was measured. As a control, 500 μL of deionized water was used instead of the sample. For the blank test, 750 μL of ethanol was used instead of DPPH and distilled water. Ascorbic acid, which is known to have antioxidative activity, was used as a positive control. The antiradical activity was also calculated as% of DPPH decolorization according to the following formula.

As a result, ascorbic acid, which was used as a positive control, showed an activity of 0.04 mg / ml for EC 50 and Chaetoceros The antioxidative activity of gracilis extracts showed a radical scavenging ability of EC 50 of 0.6 mg / ml. Therefore, it was confirmed that the microalgae extract of the present invention had a radical scavenging ability almost equivalent to that of the antioxidant used in the prior art (see Table 1).

The Chaetoceros DPPH (EC50) measurement results of gracilis extract sample EC ₅₀ , mg / mL Ascorbic acid (positive control) 0.04 Chaetoceros gracilis extract 0.6

< Experimental Example  2>

Chitoseros Gracilis ( Chaetoceros gracilis) of  Anti-inflammatory activity measurement for scalp improvement, prevention of hair loss, and therapeutic effect

In addition to radical formation, inflammation is known to be responsible for causing diseases of scalp and hair. Therefore, when the inflammation is inhibited, hair loss can be prevented and treated. Therefore, the present inventors have confirmed through experiments such as the following that the microalgae extract of the present invention has anti-inflammatory activity.

<2-1> RAW  264.7 Cytotoxicity measurement for cells

The present inventors have found that Chaetoceros Before measuring the anti-inflammatory effect of gracilis extract, we first examined the cytotoxicity of microalgae extracts on RAW 264.7 cells distributed from Korean Cell Line Bank (KCLB, Seoul, Korea).

That is, RAW 264.7 cells were cultured in Dulbecco's modified Eagle's Medium (DMEM) medium containing 10% FBS at a concentration of 1 × 10 ⁵ cells / well in a cell culture flask, At a temperature of 37 &lt; 0 &gt; C for a period of time, 95% humidity and CO _{2 ,} 5% CO ₂ When the cells were cultured in an incubator, the microalgae extract of the present invention was treated with cells to final concentrations of 0.1, 1, 10 and 100 μg / mL, respectively, and then cultured for 24 hours. The viability of the cells was measured at 490 nm using a microplate reader (Molecular Devices, VersaMax ELISA Microplate Reader, USA) using MTS reagent. Cell viability was determined by measuring the absorbance of the sample treated group Respectively.

As a result of the analysis, as shown in Fig. 1, Chaetoceros The toxicity of gracilis extract did not show toxicity to the cells up to the concentration of 200 ug / ml, but rather the cell proliferation increased to the concentration of 100 ug / ml in proportion to the treatment concentration. Therefore, the inventors of the present invention have found that the extract of the present invention does not cause specific cytotoxicity to cells.

<2-2> Nitric oxide production inhibitory activity assay

The present inventors have found that Chaetoceros To determine the anti-inflammatory effect of gracilis extract, RAW 264.7 cell line was divided into 96 wells and cultured until the cells reached about 80% of the bottom. Then, lipopolysaccharide (LPS), an inflammation inducer, was added at a concentration of 1 μg / Respectively. Then, Chaetoceros gracilis extract of the present invention was treated at 0.1, 1, 10 and 100 μg / mL, respectively. After culturing for 24 hours, the amount of nitric oxide (NO) produced was analyzed by Griess reagent. The control group was treated with lipopolysaccharide (LPS) alone or with no treatment.

The analysis using the Griess reagent was performed by first adding 1% sulfanilamide to 5% nitric acid solution, adding 0.1% naphthylethylene-diamine di-hydrochloride and mixing, and then taking 50 μL of the supernatant of the cultured cells in a 96-well plate , 50 μL of Griess reagent was added to the mixed solution, and the mixture was reacted at room temperature for 15 minutes. Absorbance was measured at 540 nm using a microplate reader (Molecular Devices, VersaMax ELISA Microplate Reader, USA). The amount of nitric oxide production was expressed as the amount of nitric oxide produced in the sample treated group based on the amount of nitric oxide produced in the control group that was not treated with the sample.

As a result of the analysis, as shown in FIG. 2 and Table 2, in the group treated with LPS alone, inflammation was induced as compared with the group treated with nothing, and the amount of nitrogen oxide production was increased, whereas in the group treated with the microalgae extract of the present invention , The amount of nitric oxide production decreased in proportion to the treatment concentration. In particular, the EC ₅₀ , mg / mL of the extract of the present invention was found to be 93.4.

The Chaetoceros NO (EC50) measurement results of gracilis extract sample EC ₅₀ , mg / mL Chaetoceros gracilis 93.4

<2-3> PGE2  Production inhibition activity assay

It is known that prostaglandin (PGE) is an immunomodulator and has an activity of inducing an inflammatory reaction. Therefore, when the production or activity of the prostaglandin is inhibited, the inflammatory reaction can be inhibited, thereby preventing hair loss due to inflammation. The present inventors have found that Chaetoceros gracilis extract of the present invention was tested for inhibition of PGE ₂ production using PGE ₂ extract of the present invention using Prostaglandin E ₂ EIA Kit (Cayman Chemical Company; Ann Arbor, Michigan, USA) Production inhibitory effect. At this time, the measurement method was quantified according to the analysis method of the manufacturer. First, RAW 264.7 cells were treated with Chaetoceros gracilis extracts were treated with 0.1% LPS at concentrations of 0.01, 0.1, 1, 10, 100, and 200 ug / mL for 24 hours, respectively. Cell culture supernatants were then added to a 96-well plate coated with goat anti-mouse 50 μL of the primary antibody solution and 50 μL of PGE ₂ conjugate were added thereto, followed by reaction at 4 ° C. for 18 hours, followed by washing with washing solution 5 times, followed by 200 μL of Ellman's reagent, stirring for 60 minutes After the reaction, the absorbance was measured at 405-420 nm.

As shown in FIG. 3, the amount of PGE ₂ produced by stimulation with LPS was inhibited by the microalgae extract of the present invention, and the degree of inhibition of PGE ₂ production was inhibited . Especially, the group treated with the extract at a concentration of 100 ug / mL inhibited the production of PGE ₂ by 37.7%.

< Experimental Example  3>

Chitoseros Gracilis ( Chaetoceros gracilis) of  5 alpha for scalp improvement and hair loss prevention effect Reductase  activation Low performance  analysis

The 5 alpha redutase enzyme is known to have an activity to increase the activity of testosterone, a male hormone that interferes with the production of energy required for hair growth and development. Therefore, a substance which inhibits the activity of 5 alpha reductase is used as a hair loss prevention and hair growth promoting agent.

Therefore, the present inventors have found that Chaetoceros gracilis to extract the Confirm has the 5-alpha-reductase activity inhibition, SD rat liver a 0.32 M sucrose - 1 mM dithiothreitol / 0.02 M phosphate buffer was placed in a (pH 6.5) Homogenization was separated and then centrifuged 30 minutes (4500 × g , 0 C). After that, the supernatant was measured for protein by BCA method, and the concentration was adjusted to 5.0 mg / mL and stored at -50 ° C. The activity inhibition of 5 alpha-reductase was measured by modifying the method of Kumar et al. (2011). (100 μL), testosterone / 50% ethanol (30 μL), rat microsoma supernatant (100 μL), and 0.77 mg / mL NADPH 0.5 uL was added thereto, followed by reaction at 37 ° C for 30 minutes. The reaction was stopped by adding 500 μL of dichloromethane and 5.0 mL of dichloromethane, and 50 μL of 100 ppm propyl p-hydroxybenzoate / 50% ethanol (as an internal standard for HPLC) was added. The mixture was centrifuged for 10 minutes to obtain a dichloromethane layer. The dichloromethane layer was concentrated with nitrogen and then dissolved in 250 uL of methanol. For the determination of remaining testosterone, 10 μL of the reaction stop solution was injected into the HPLC equipped with a C ₁₈ reverse phase column (Bondclone C ₁₈ , 5 μm, 250 × 4.6 mm, Daiso Co., Ltd. Japan), and 65% methanol solution Was detected at 245 nm with elution at 1 mL / min. At this time, the temperature of the column was adjusted to 40 ° C.

As a result of the analysis, as shown in Table 3 and FIG. 4, in the case of Finasteride, which is an FDA approved drug, the IC50 was 0,782 ug / ml as a result of inhibition of 5-alpha reductase enzyme activity, and Cheatoceros gracilis The inhibitory activity of the extract was 7.5 ± 2.1% at 0.1 mg / ml, 46.6 ± 3.5% at 1 mg / ml and 65.1 ± 2.3% at the concentration of 10 mg / ml.

In addition, the present invention is significant in that it is an extract which is not a high activity but is not a single substance compared to the pinastelide, and that a material having anti-hair-removing effect is extracted from a material derived from a natural material. Therefore, It will be more useful in the future.

The Chaetoceros Inhibitory activity of 5-alpha-reductase activity of gracilis extract Sample, mg / ml Inhibition activity,% Chaetoceros gracilis 0.1 7.5 ± 2.1% One 46.6 ± 3.5% 10 65.1 ± 2.3% Finasteride IC ₅₀ 0.782 ug / mL

The present invention has been described with reference to the preferred embodiments and experimental examples. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (8)

A pharmaceutical composition for improving scalp and preventing hair loss, which contains an extract of Chaetoceros gracilis, which is a microalgae, as an active ingredient. The method according to claim 1,
Wherein the extract is an extract extracted with water or an organic solvent. The method according to claim 1,
Wherein the extract comprises Chaetoceros gracilis microalgae, a culture of the microalgae, a concentrate of the culture, or a dried product of the culture. The method according to claim 1,
Wherein said extract has antioxidant activity, anti-inflammatory activity and inhibitory activity against 5-alpha-reductase enzyme activity. The method according to claim 1,
Wherein the pharmaceutical composition is selected from the group consisting of a cream, a gel, a patch, a spray, an ointment, a warning agent, a lotion, a liniment, a pasta, and a cataplasma as an external preparation for skin. A pharmaceutical composition. A health functional food for improving the scalp and preventing hair loss, which contains an extract of Chaetoceros gracilis, which is a microalgae, as an active ingredient. A cosmetic composition for improving the scalp and preventing hair loss, which comprises an extract of Chaetoceros gracilis, which is a microalgae, as an active ingredient. 8. The method of claim 7,
Wherein the cosmetic composition is any one selected from the group consisting of hair tonic, hair nourishing lotion, scarf treatment, hair treatment, hair rinse, hair shampoo and hair lotion.

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