KR20140020005A - Cistanche tubulosa, fucoidan extract for promoting hair growth and preventing hair loss & manufacturing method thereof - Google Patents

Abstract

The present invention relates to a Cistanche tubulosa and fucoidan extract having effects of preventing hair loss and promoting hair growth and a manufacturing method of the same and more specifically, to a Cistanche tubulosa and fucoidan extract for preventing hair loss and promoting hair growth and a manufacturing method of the same wherein the extract contains at least one extract as an active ingredient among: an extract, as a phenylethanoid glycoside, including at least one among echinacoside, acteoside, and a sulfuric acid group; and an extract including at least one among the phenylethanoid and the sulfuric acid group. [Reference numerals] (AA) Seaweed(Undaria pinnatifida sporophyll/Laminaria japonica); (BB) Pulverizing; (CC) Extract acid and hot water; (DD) Separate solid-liquid; (EE) Supernatant neutralization; (FF) Diatom filtration; (GG) Ultrafilter; (HH) Alcohol immersion; (II) Centrifugation; (JJ) Drying; (KK) Packaging; (LL) Alginic acid separation by depositing; (MM) 10N NaOH solution; (NN) Three times capacity of ethanol is added

Description

Extract and method for producing sarcoma and fucoidan extract having hair loss prevention and hair growth promoting effect {Cistanche Tubulosa, Fucoidan Extract for Promoting Hair Growth and Preventing hair loss & Manufacturing method Thereof}

The present invention relates to a breeding and fucoidan extract having a hair loss prevention and hair growth promoting effect comprising at least one of the extract for breeding and fucoidan extract as an active ingredient and a method for producing the same.

In recent years, hair loss is increasing not only in middle age but also in adolescence, which is causing a lot of problems in social activities. Although hair abnormalities do not affect life, they often cause great difficulties in interpersonal relationships caused by mental stress. Therefore, much attention is focused on the search for a safe and easy material that can prevent hair loss, promote regrowth of hair, and promote hair growth. Current effective hair repellents include minoxidil (MXD), finasteride and propecia. MXD was developed as a vasodilator for the treatment of hypertension, but as a side effect, it was developed as a hair restorer (Hurton and Marshall, 1979). The mechanism of action on the hair growth effect of minoxidil has not been clarified until now, but it is thought that nutrient expansion through potassium expansion and potassium channel opening effect induce hair growth (Meisheri et al., 1988; Buhl et al. , 1990a, 1990b). MXD has been shown to be effective against male-pattern baldness, orrogenic alopecia caused by male hormones, even among the baldness treatments. Finasteride and propecia, on the other hand, are agents for the treatment of prostatic hyperplasia and prostate gland cancer, and they inhibit type II 5-reductase, an enzyme that converts testosterone to dihydrotestosteron (DHT), the causative agent of prostatic hypertrophy and hair loss (Kaufman et al., 1999). This enzyme is present in the hair follicle cells at the base of hair follicles. Finasteride and propecia are effective for male hair loss.

Hair follicles themselves are remoteled through cycles of anagen, catagen, telogen and exogen (Paus and Cotsarelis, 1993). During catagen, hair follicles undergo a programmed cell death (apoptosis) process, downsizing into telogen (Cotsarelis, 1997). Hair follicles regenerate at the beginning of the next anagen phase, requiring activation of stem cells in the swelling of the hair follicles (Cotsarelis et al., 1990). Proliferated stem cells form new hair follicle matrix during the early anagen phase, and the hair shaft and inner root sheath are derived from relatively undifferentiated basal cells (Oshima et al., 2001). The size and length of the hair stem are proportional to hair follicle size and anagen duration, respectively. The periodic growth of these coats is regulated by various growth factors.

The most common forms of alopecia are classified as masculine alopecia or general baldness, telogen effluvium, anticancer drug-induced alopecia and alopecia areata. C57BL / 6 C3H / HeJ mice are used as model animals for hair loss studies. C57BL / 6 mice are characterized by black hair, spontaneous alopecia, and limited melanocytes in hair follicles, and the synthesis of melanin coincides well with the hair growth cycle. As a result, it is widely used in hair growth research because it has the advantage of determining the hair growth cycle (Pals et al., 1989; Mller-Rver et al., 2001). C3H / HeJ mice have similar histological and immunohistochemical characteristics to human alopecia areata (Sundberg et al., 1994). Female C3H / HeJ mice over 6 months develop alopecia areata at a frequency of 20%, in which lymphocytes infiltrate around the hair follicles and steroid agents are effective in the lesions, thus resembling immunotherapy in humans (Freyschmidt-Paul et al. , 1999).

Sulfate polysaccharide complex of fucoidan derived from seaweed, such as kelp (Laminaria japonica), miyeokgwi (Undaria pinnatifida Sporophyll), Mozuku (Cladosiphon okamuranus) distributed in many countries has long been used as a remedy in oriental medicine. In previous studies, fucoidan has been reported to have antioxidant, anticoagulant, anticancer and anti-inflammatory effects (Feldman et al., 1999; Wang et al., 2010). Therefore, the usefulness of fucoidan for inflammatory diseases, ischemia, immune suppression and tumors has attracted researchers' attention (Bojakowski et al., 2001; Li et al., 2005). In recent years, fucoidan has been reported to have excellent thrombolytic effect, thereby preventing and improving ischemic disease (Min et al., 2012a, 2012b).

Meanwhile, breeding ( Cistanche tubulosa ) has traditionally been widely used in China. Sarcoma use has been reported to reduce the production of TNF- and IL-4, the major cytokinase required for NO release from inflammatory cells, and showed strong anti-inflammatory effects in animal experiments (Yamada et al., 2010; Kyung et al., 2012). . Sarcoma is also known to exert a blood circulation improvement effect by lowering blood cholesterol (Shimoda et al., 2009).

The technical problem to be solved by the present invention is to find a material that has safe and effective hair loss prevention and hair growth promoting activity from natural products, seaweeds such as kelp (Laminaria japonica), Undaria pinnatifida Sporophyll, Cladosiphon okamuranus Check whether the complex prescription of fucoidan and sarcoma extract, a sulfate polysaccharide complex, is promoted hair growth, and not only the existing shampoo or hair tonic form applied to the scalp, but also the health function Food and medicine, but also for breeding and fucoidan extract having a hair loss prevention and hair growth promoting effect that can maximize the hair loss prevention and hair growth effect and a method for producing the same.

In order to achieve the above object, the breeding and fucoidan extracts having the hair loss prevention and hair growth promoting effect of the present invention is a phenyl ethanoid glycoside, containing at least any one of the equinacoside, acteoside and sulfate group and phenyl Characterized in that it comprises at least one extract of the extract including at least one of the tanoid glycosides and sulfate groups as an active ingredient.

At this time, the extract containing at least any one of the equinacoside, acteoside and sulfate group as the phenyl ethanoid glycosides is an extract for breeding which is a heat sugar and a plant, the extract containing at least one of the phenyl ethanoid glycosides and sulfate groups It is characterized in that the fucoidan extract.

In addition, the composition is characterized by having a -Glutamyl transpeptidase (-GTP) activity.

In addition, the composition is characterized by having alkaline phosphatase (ALP) activity.

In addition, the composition is characterized in that it has an activity in the expression amount of Epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF).

On the other hand, as a phenyl ethanoid glycoside, the extract containing at least one of echinacoside, acteoside and sulfate group is 10 ~ 500 mg / body, the extract containing at least one of the phenyl ethanoid glycoside and sulfate group is 100 ~ It is characterized by using 2,000mg / body.

In addition, the extract for the breeding, extraction step for extracting the breeding with water or ethanol; Filtration step; Solvent removal step of removing the solvent water or ethanol using a vacuum evaporator; Partial purification step of partial purification through column chromatography; Active substance generation step of obtaining an active substance of 25% or more of echinacoside and 9% or more of acteoside by hot air drying; And a particle selection step of selecting fine particles using a sieve of 50 mesh.

In addition, the fucoidan extract, the grinding step of grinding the fucoidan raw material; Extraction step of acid and hot water extraction for 2 hours at pH 2.0, 60 ℃; Precipitating separation step of precipitating alginic acid by solid-liquid separation; Neutralizing the supernatant with 10N NaOH; Filtration step of diafiltration and ultrafiltration; A precipitation step of precipitating with ethanol three times the volume; Centrifugation step; And characterized in that it comprises a drying step.

According to the present invention described above, by using at least one of the extract for breeding and fucoidan extract can prevent hair loss and hair growth promoting effect.

In addition, the sarcoma extract and fucoidan extract can be used in hair care products such as shampoo or hair tonic applied to the scalp, as well as health functional foods and pharmaceutical hair oral compositions.

1 is a view showing the hair growth analysis over time on day 14 of the extracts for fucoidan and sarcoma according to one embodiment of the breeding and fucoidan extract having hair loss prevention and hair growth promoting effect according to the present invention,
2 is a view showing the hair growth analysis over time on day 21 of the extracts for fucoidan and sarcoma according to one embodiment of the breeding and fucoidan extract having hair loss prevention and hair growth promoting effect according to the present invention,
3 is a graph showing the effect of increasing the hair growth rate of 0 to 21 days of fucoidan and sarcoma extract according to one embodiment of the breeding and fucoidan extract having hair loss prevention and hair growth promoting effect according to the present invention,
4 is an oral administration of fucoidan and sarcoma extract according to one embodiment of the breeding and fucoidan extract having hair loss prevention and hair growth promoting effect according to the present invention, the 14th, a view showing histological changes,
5 is an oral administration of fucoidan and sarcoma extract according to one embodiment of the breeding and fucoidan extract having hair loss prevention and hair growth promoting effect according to the present invention, on the 21st, a diagram showing a histological change,
Figure 6 is a diagram showing the -Glutamyl transpeptidase (-GTP) activity after oral ingestion according to one embodiment of the breeding and fucoidan extract having hair loss prevention and hair growth promoting effect according to the present invention,
7 is a view showing alkaline phosphatase (ALP) activity after oral ingestion according to one embodiment of the breeding and fucoidan extract having hair loss prevention and hair growth promoting effect according to the present invention,
8 is a view showing the measurement of Epidermal growth factor (EGF) expression after oral ingestion according to one embodiment of the breeding and fucoidan extract having hair loss prevention and hair growth promoting effect according to the present invention,
9 is a view showing the measurement of vascular endothelial growth factor (VEGF) expression after oral ingestion according to one embodiment of the breeding and fucoidan extract having hair loss prevention and hair growth promoting effect according to the present invention,
10 is a view showing the extraction and manufacturing process for breeding according to one embodiment of the method for producing a breeding and fucoidan extract having hair loss prevention and hair growth promoting effect according to the present invention,
11 is a view showing a fucoidan extraction and manufacturing process according to an embodiment of the method for producing a breeding and fucoidan extract having a hair loss prevention and hair growth promoting effect according to the present invention.

While the invention is susceptible to various modifications and alternative forms, specific embodiments thereof are shown by way of example in the drawings and will herein be described in detail.

It should be understood, however, that the invention is not intended to be limited to the particular embodiments, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention. Like reference numerals are used for like elements in describing each drawing.

The terms including ordinal, such as second, first, etc., may be used to describe various elements, but the elements are not limited to these terms.

The terms are used only for the purpose of distinguishing one component from another. For example, without departing from the scope of the present invention, the second component may be referred to as a first component, and similarly, the first component may also be referred to as a second component. And / or < / RTI > includes any combination of a plurality of related listed items or any of a plurality of related listed items.

When a component is referred to as being "connected" or "connected" to another component, it may be directly connected to or connected to that other component, but it may be understood that other components may be present in between. Should be. On the other hand, when an element is referred to as being "directly connected" or "directly connected" to another element, it should be understood that there are no other elements in between.

The terminology used in this application is used only to describe a specific embodiment and is not intended to limit the invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this application, the terms "comprise" or "have" are intended to indicate that there is a feature, number, step, operation, component, part, or combination thereof described in the specification, and one or more other features. It is to be understood that the present invention does not exclude the possibility of the presence or the addition of numbers, steps, operations, components, components, or a combination thereof.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries are to be interpreted as having a meaning consistent with the contextual meaning of the related art and are to be interpreted as either ideal or overly formal in the sense of the present application Do not.

The present invention, as a result of the effort to find a safe and effective hair loss prevention and hair growth promoting effect from natural products, at least one of the extract for breeding, fucoidan extract experimentally confirmed that the hair loss prevention and hair growth promoting effect is excellent It was completed.

Meanwhile, low-dose fucoidan (18 mg / kg) and sarcoma extract (54 mg / kg) were converted to body surface area-to-weight ratios (adult Km = adult) from the estimated clinical doses of 100 mg / body (70 kg) and 300 mg / body, respectively. 37 / mouse Km = 3).

The rationale for establishing the expected dose is as follows. According to a clinical trial conducted by the Chinese Pharmaceutical Industry Co., Ltd. (Hangzhou) Pharmaceutical Co., Ltd., the basis for the use of total glycosides (including echinaceoside and acteoside) for breeding as an anti-vascular dementia drug In phase I clinical trials for sarcoma, no adverse effects and adverse changes in each observation parameter were observed during single and continuous dose safety trials, so that the normal population had good water resistance against total glycoside capsules for sarcoma. In clinical trials, it became clear that it was safe to use in clinical trials. In Phase II clinical trials, 600 mg or 3 times a day were given orally with respect to the dosage and dosage. As a result, the clinical results of five facilities using randomization, double-blind, double pile parallel test, and multi-center test method were assured against the treatment of vascular dementia (deficiency deficiency) of total glycoside capsules for breeding. Since no serious adverse events such as drug toxicity and side effects occur during clinical trials and clinical trials, it has been clarified that total glycoside capsules for sarcoma are effective and safe drugs for the treatment of vascular dementia. The clinical results of phase III were also similarly effective in the treatment of vascular dementia (lack of hydrolysis) of total glycoside capsules for sarcoma based on the results of clinical trials at four facilities using randomization, parallel testing, and multi-center test methods. During the clinical trials, no serious adverse events such as toxicity and side effects by the drug occurred, and the total glycoside capsule for sarcoma proved to be an effective and safe drug for treating vascular dementia.

The term extract used in the present invention while referring to a fucoidan extract which is a sulfate polysaccharide complex obtained from seaweeds such as extracts for breeding, kelp (Laminaria japonica), seaweed (Undaria pinnatifida Sporophyll), and seaweed such as Cladosiphon okamuranus. Silver is meant to include not only the extraction result obtained by treating the extraction solvent, but also for breeding and fucoidan processed products formulated to administer the algae itself, such as breeding and kelp, seaweed, and large horsetail, to the subject.

The MK-R7 extract of the present invention may be separated according to conventional methods known in the art for extracting extracts from natural products, that is, using conventional solvents under the conditions of conventional temperature, pressure.

MK-R7 used in the composition of the present invention can be used a variety of extraction solvent when obtained by treating the extraction solvent.

Preferably, polar solvents or nonpolar solvents may be used, and suitable as polar solvents are water, alcohols (preferably methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, 1-pentanol). , 2-butoxyethanol or ethylene glycol, acetic acid, dimethyl-formamide (DMFO) and dimethyl sulfoxide (DMSO).

Suitable as nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- Pentene, 1-chlorobutane, 1-chloropentane, 0-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloro Methane, 1,2-dichloroethane, anneal, diethylamine, ether, carbon tetrachloride and THF (tetrahydrofuran).

More preferably, the extraction solvent used in the present invention is water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), a mixed solvent of the lower alcohol and water, acetone, ethyl acetate , Chloroform, butyl acetate, 1,3-butylene glycol, hexane and diethyl ether. Most preferably, the extract of the present invention is obtained by treating MK-R7 with water or ethanol.

Including the term active ingredient in the present invention means that the MK-R7 extract contains an amount sufficient to achieve efficacy or activity.

The present invention is an extract extracted from the natural plant material for breeding and kelp, seaweed, large horsetail, etc., even if excessive administration, so there is no side effect to the human body, the upper limit of the quantity of breeding and fucoidan extract can be carried out by those skilled in the art to select within an appropriate range.

Suitable dosages of the pharmaceutical compositions of the present invention may vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion and reaction sensitivity of the patient, Typically, the skilled practitioner can readily determine and prescribe a dosage effective for the desired treatment or prophylaxis.

The daily dosage of the pharmaceutical composition of the present invention is 0.0010,000 mg / kg.

The pharmaceutical composition of the present invention may be prepared in unit dose form or prepared by incorporation into a multi-dose container by formulating with an excipient according to a method which can be easily carried out by one of ordinary skill in the art. Can be.

Here, the formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The extract of the present invention may be provided in the form of a functional food composition for improving scalp disease by hair loss prevention and hair growth promoting effect.

Functional foods, quasi-drugs, and pharmaceutical compositions of the present invention include ingredients that are commonly added during food preparation, and include, for example, proteins, carbohydrates, fats, nutrients, and seasonings.

For example, when prepared with a drink, flavoring agents or natural carbohydrates may be included as additional ingredients in addition to the active ingredient.

In addition, natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.); Disaccharides (e.g., maltose, sucrose, etc.); oligosaccharide; Polysaccharides (e.g., dextrin, cyclodextrin and the like); And sugar alcohols (e.g., xylitol, sorbitol, erythritol, etc.).

As the flavoring agent, natural flavoring agents (e.g., taumartin, stevia extract, etc.) and synthetic flavoring agents (e.g., saccharin, aspartame, etc.) can be used.

The extract of the present invention may be provided in the form of a cosmetic composition for improving skin diseases.

Cosmetic compositions of the present invention can be prepared in any formulations conventionally prepared in the art and include, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing It may be formulated as cleansing, oil, powder foundation, emulsion foundation, wax foundation, spray, and the like, but is not limited thereto.

More specifically, it may be prepared as a flexible lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing cream, a foam, a cleansing water, a pack, a patch, a spray or a powder formulation.

Extracts of the present invention can be used by methods such as direct application or scattering on the hair or scalp. The hair to which the extract of the present invention is applied includes all parts of the hair root and hair follicles, such as hair roots and hair follicles of the head, and overall hair.

Therefore, the extract can be used in hair tonic, hair lotion, hair cream, hair spray, hair mousse, hair gel, hair conditioner, hair shampoo, hair rinse, hair pack, hair treatment and the like.

In addition, the plant with the heat sugar family used as a raw material is not specifically limited, For example, all parasitic plants (Cistanche tubulosa: for mandarin breeding), cistanche salsa (for breeding), herbicide (Aeginetia indica), alder tree (Boschniakia) rossica), but the present invention is not limited thereto.

In addition, only 1 type may be used, 2 or more types may be used together, and most preferably, a parasitic plant (Cistanche tubulosa: for mandarin sarcoma) is used.

On the other hand, the preferred complex extract of the present invention is fucoidan 18 or 54 mg / kg, breeding extract 54 or 162 mg / kg, or a mixture thereof 18 + 54, 54 + 162 or 90 + 27 mg / kg, of the present invention The most preferred composition is synergistic effect of promoting hair growth when combined with a high dose (54 + 162 mg / kg, clinical dose 300 mg / body (70 kg) + 900 mg / body) of the sarcoma extract and fucoidan.

Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings.

1. Test substance

For breeding, as shown in FIG. 11, extraction with water or ethanol → filtration → removal of water and ethanol as solvents using a vacuum evaporator → partial purification through column chromatography → hot air drying to more than 25% echinacoside, more than 9% acteoside Obtaining active substance → Selecting fine particle powder using a sieve of 50 mesh → Commercialization

Fucoidan, as shown in Figure 12, pulverized fucoidan raw material (Dashima) → acid / hot water extraction for 2 hours at pH 2.0, 60 ℃ → separation of alginic acid precipitation by solid-liquid separation → neutralization of the supernatant with 10N NaOH → diatom filtration and ultrafiltration Filtration → precipitation with three volumes of ethanol followed by centrifugation and drying.

The test substance was stored at 4 ° C, and then fucoidan: breeding extract 1: 3 was mixed before use, dissolved in purified water and orally administered at a dosage of 5 ml / kg (0.1 ml per animal = 100 μl orally). Kyung et al., 2012). [LAR: Laboratory Animal Research-Lab Anim Res 2012: 28 (2)]

2. Experimental animals

Five-week-old male C57BL / 6 mice were supplied from Orient Bio (Gapyeong, Korea) and used for about one week after acclimation. Animals were housed 4 in a mouse cage (220 × 200 × 145 mm). The environment of the animal laboratory was adjusted to a temperature of 23 ± 2 ° C, a relative humidity of 55 ± 10%, a ventilation frequency of 12 times / hour, a lighting cycle of 12 hours, and an illuminance of 150-300 Lux. Purina Rat Chow ^ⓡ , a pellet-type solid food for experimental animals, was supplied from ^Biopia (Gunpo, Korea).

3. Administration of test substance

At 6 weeks of age, the dorsal area of the mouse (n = 8 / group) was epilated with an electric razor to induce the telogen hair roots into the anagen (Hattori and Ogawa, 1983).

That is, wait until the hair is uniformly pink, and when pink is confirmed, the remaining hair is lightly shaved so as not to damage the skin with the razor (Lee et al., 2006; Kim et al., 2009).

From the day after epilation, fucoidan (18 or 54 mg / kg), sarcoma extract (54 or 162 mg / kg), or a mixture thereof (18 + 54, 54 + 162 or 90 + 270 mg / kg) is administered orally daily for 21 days. It was.

Low-dose fucoidan (18 mg / kg) and sarcoma extract (54 mg / kg) were converted to body surface area-to-weight ratio (adult Km = 37 /) from the estimated dead dose of 100 mg / body (70 kg) and 300 mg / body, respectively. It calculated by applying mouse Km = 3).

High doses were administered three times as low doses alone or in combination, and the highest dose was set at five times to evaluate the highest synergistic effect between the two test substances (Kyung et al., 2012). MXD was applied to the hair removal site by using a brush at an amount of about 0.2 mL once daily for 21 days from the day after hair removal.

4. Hair Growth Analysis Over Time

The dorsal area was photographed to determine the hair growth effect before (0 day) and 3 days after administration of the test substance. Using a computerized image analysis system (Imageinside, Daejeon, Korea), the ratio of the hair growth area (regrowth area / shaved area) per unit of skin epilation was obtained.

4-1. Hair growth analysis results over time (see FIGS. 1 to 3)

When the MXD (3%) was applied to the skin, a faster hair growth promoting effect was confirmed on the 14th day (FIG. 1B and FIG. 3) compared to the control group (FIG. 1A and FIG. 3).

In addition, even on day 21, the MXD-coated group (B and FIG. 3 of FIG. 2) had much more hair growth than the control group (A and FIG. 2 of FIG. 2).

In particular, about 15 days after application, MXD occupied 62% of the skin, but only 6% of the hair was found in the control group. Even at 21 days, it was 93% in the MXD-coated group compared to about 30% in the control group.

In contrast, the Fucoidan oral administration group (C, D in FIG. 1, C and D in FIG. 1) was lower than the MXD-coated group, but was significantly promoted in the high dose (54 mg / kg) group compared to the control group by 69%. Reached (FIG. 3).

The sarcoma extract also promoted hair growth up to 74% at 21 days, similar to fucoidan 54 mg / kg at 54 mg / kg (E in FIG. 1, E in FIG. 2 and FIG. 3), and at 21 days at high doses of 162 mg / kg. To 87% (F in FIG. 1, E in FIG. 2 and FIG. 3).

On the other hand, synergistic effects were not observed when low dose fucoidan and sarcoma extract (18 + 54 mg / kg) were administered (G of FIG. 1, G of FIG. 2 and FIG. 3), but high dose (54 + 162 mg / kg). ), Synergistic effect was observed, and the superior promoting effect was confirmed from the 12th day of administration compared to the control group, and increased to 46% (control group 6%) on the 15th day and 86% (control group 30%) on the 21st day. (H of FIG. 1, H of FIG. 2, and FIG. 3), it confirmed that it was comparable to MXD.

However, at the highest dose (90 + 270 mg / kg) of the combined dose no effect was increased (I in Figure 1, I in Figure 2 and Figure 3).

5. Histological examination

On the 14th and 21st day after the start of the test, the hairs were extracted to include the areas of strong hair growth and delayed areas, and fixed with 10% neutral formalin solution.

After embedding the tissue in paraffin, 5 tissue sections were prepared and stained with hematoxylin-eosin to observe the number and histological changes of hair follicles under an optical microscope.

5-1. Histological examination results (see FIGS. 4 and 5)

Fidelity of the hair root was significantly improved in the skin histological findings on day 14 and day 21 of MXD application compared to the control group (A of FIG. 4 and A of FIG. 5) (FIG. 4B and FIG. 5B).

This phenomenon was also observed in the fucoidan-administered group, the sarcoma extract-administered group, and their combined administration group (C-4 I of FIG. 4 and C-5 I of FIG. 5).

In particular, the most effective in the Fucoidan 54 mg / kg + breeding extract 162 mg / kg complex prescription group was confirmed to be superior to the MXD application group.

6. Measurement of -Glutamyl transpeptidase (-GTP) and alkaline phosphatase (ALP) activities

The extracted skin tissues were immediately frozen in liquid nitrogen, and 4 times of phosphate buffer (pH7.4) was added and homogenized.

The homogenate was centrifuged at 3,000 rpm for 20 minutes, and the supernatant was used for -GTP and ALP activity measurements.

-GTP and ALP activities were measured by enzyme rate assay (GSCC) using P -nitrophenolphosphate (PNPP) -DEAV (Handiiski et al., 1994; Kim et al., 2009).

6-1. -Glutamyl transpeptidase (-GTP) and alkaline phosphatase (ALP) activity measurement results

As a result of measuring -GTP activity, which is known to induce hair root into anagen, the MXD-applied group increased about 120% on day 14 and about 55% on day 21 compared to the control group (FIG. 6). Oral administration of fucoidan also showed a tendency to increase -GTP activity at high doses (54 mg / kg).

In particular, in the high dose (162 mg / kg) sarcoma extract administration group was found a significant increase on day 14.

Interestingly, when combined administration of fucoidan and sarcoma extract showed a synergistic effect compared to each administration group.

Nevertheless, there was no synergistic effect in the combined dose group of the highest dose (fucoidan 90 + 270 mg / kg breeding extract).

In addition, the ALP activity, known as an indicator of skeletal growth and angiogenesis in animals, was significantly increased in the MXD-coated group at 14 and 21 days compared to the control group (FIG. 7).

Fucoidan and sarcoma extracts also showed high deviations but increased ALP activity.

Remarkably, the combination of fucoidan and sarcoma extract was higher than the administration of each of them.

7.Epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) expression levels

EGF and VEGF were measured using enzyme-linked immunosorbent assay (ELISA) kits (R & D Systems, Minneapolis, USA) (Cross et al., 2003; Kim et al., 2009).

That is, according to the manufacturer's instructions, the production of two growth factors was quantified by sandwich enzyme immunoassay using polyclonal antibodies specific for mouse EGF and VEGF.

7-1. Epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) expression levels

As a result of analyzing EGF promoting hair growth, it was increased by more than 3.5 times in 14 and 21 days in the MXD group compared to the control group (FIG. 8).

In comparison, fucoidan induced a slight increase on day 14, but showed a significant increase on day 21.

On the other hand, the extract for breeding was weak at low dose (54 mg / kg), but showed an excellent effect at high dose (162 mg / kg).

Significantly, when combined with fucoidan and sarcoma extract, the synergistic effect between the two substances was confirmed by a significant increase in EGF content.

The angiogenic factor VEGF was analyzed to increase about 80% on day 14 and about 50% on day 21 in the MXD group compared to the control group (FIG. 9).

In comparison, fucoidan had no effect, but the sarcoma extract showed a significant VEGF synergistic effect.

In addition, the synergistic effect of VEGF was increased on the 14th day when a combination of fucoidan and sarcoma extract was administered.

8. Statistical Processing

All measurements are expressed as mean standard error. When significance was observed after one-way ANOVA analysis, Turkey's test was used to test the significance between groups at the level of P <0.05.

Claims (10)

Hair loss prevention using as an active ingredient an extract containing at least one of echinaceoside, acteoside and sulfate group as an phenylethanoid glycoside and an extract containing at least one of phenylethanoid glycoside and sulfate group And extracts for breeding and fucoidan having hair growth promoting effect.
The method of claim 1,
As the phenyl ethanoid glycoside, the extract containing at least one of echinaceoside, acteoside and sulfate group is an extract of a heat sugar and a plant, and the extract containing at least one of the phenyl ethanoid glycoside and a sulfate group is a fucoidan extract. For breeding and fucoidan extract having hair loss prevention and hair growth promoting effect, characterized in that.
3. The method of claim 2,
The heat sugar and plants are any one or more of breeding, and fucoidan is a fucoidan, a sulfate polysaccharide complex (sulfate polysaccharide complex) obtained from seaweed, breeding and fucoidan extract having a hair loss prevention and hair growth promoting effect.
4. The method according to any one of claims 1 to 3,
The composition is a breeding and fucoidan extract having hair loss prevention and hair growth promoting effect, characterized in that -Glutamyl transpeptidase (-GTP) activity.
4. The method according to any one of claims 1 to 3,
The composition is for breeding and fucoidan extract having a hair loss prevention and hair growth promoting effect, characterized by having alkaline phosphatase (ALP) activity.
4. The method according to any one of claims 1 to 3,
The composition for breeding and fucoidan extract having a hair loss prevention and hair growth promoting effect, characterized in that it has an activity in the amount of Epidermal growth factor (EGF) expression.
4. The method according to any one of claims 1 to 3,
The composition is for breeding and fucoidan extract having hair loss prevention and hair growth promoting effect, characterized in that it has an activity in the expression level of vascular endothelial growth factor (VEGF).
4. The method according to any one of claims 1 to 3,
As the phenyl ethanoid glycoside, the extract containing at least one of echinaceoside, acteoside and sulfate group is 10-500 mg / body, and the extract containing at least one of the phenyl ethanoid glycoside and sulfate group is 100-2,000 mg / For breeding and fucoidan extract having a hair loss prevention and hair growth promoting effect, characterized by using the body.
To prevent hair loss and hair growth promoting effect for breeding and fucoidan extract,
The extract for breeding,
Extraction step of extracting the breeding with water or ethanol;
Filtration step;
Solvent removal step of removing the solvent water or ethanol using a vacuum evaporator;
Partial purification step of partial purification through column chromatography;
Active substance generation step of obtaining an active substance of 25% or more of echinacoside and 9% or more of acteoside by hot air drying; And
A method for preparing breeding and fucoidan extracts having hair loss prevention and hair growth promoting effect, comprising the step of selecting the fine particles using a sieve of 50 mesh.
To prevent hair loss and hair growth promoting effect for breeding and fucoidan extract,
The fucoidan extract,
Grinding step of grinding the fucoidan raw material;
Extraction step of acid and hot water extraction for 2 hours at pH 2.0, 60 ℃;
Precipitating separation step of precipitating alginic acid by solid-liquid separation;
Neutralizing the supernatant with 10N NaOH;
Filtration step of diafiltration and ultrafiltration;
A precipitation step of precipitating with three times the volume of ethanol;
Centrifugation step; And
A method for producing sarcoma and fucoidan extract having a hair loss prevention and hair growth promoting effect, characterized in that it comprises a drying step.

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