KR101721685B1

KR101721685B1 - Composition for Removing dandruff, Preventing hair loss, and Promoting hair growth

Info

Publication number: KR101721685B1
Application number: KR1020150068969A
Authority: KR
Inventors: 이성표
Original assignee: (주)애니닥터헬스케어
Priority date: 2015-05-18
Filing date: 2015-05-18
Publication date: 2017-03-30
Also published as: KR20160135532A

Abstract

The present invention relates to a composition for improving dandruff removal and hair loss prevention and for promoting hair growth, which comprises a cystatin tu tubulosa extract, a seaweed extract, a broccoli sprout extract, a variety extract, an alfalfa extract, a collavi extract, 0.2): 1 (占 0): 0.5 占 0.07: 0.1 占 0.03: 0.1 占 0.03: 0.1 占 0.03: 0.1 占 0.03.

Description

{Composition for Removing dandruff, Preventing hair loss, and Promoting hair growth}

The present invention relates to a composition for prevention of hair loss and hair growth, and more particularly, to a composition for preventing hair loss and hair growth, and more particularly, to a composition for preventing hair loss and hair growth, which comprises dysentery removal including alopecia cisterna, kelp extract, broccoli sprout extract, various extract, alfalfa extract, And to a composition for promoting hair growth.

Recently, hair loss has increased not only in middle age but also in youth. Therefore, there is great interest in finding safe and easy materials that can prevent hair loss, promote regrowth of hair follicles, and promote hair growth. Currently, there are many studies on hair loss treatment, but there is no innovative development yet (Hwang, 2007). Thus, it has been scientifically proven that it is more effective to apply various combination therapies than to rely on one type of treatment.

Currently on the market are minoxidil (MXD), finasteride, propecia, and the like. Minoxidil has been developed as a vasodilator for the treatment of hypertension, but as a side effect, it has been developed as a hair growth agent (Burton and Marshall, 1979). Although the mechanism of action of the minoxidil on the hair growth effect has not been clarified to date, it is thought that the increase of nutritional supply through vasodilation and the tassium channel opening effect induce hair growth (Meisheri et al. , 1988; Buhl et al., 1990a, 1990b). These minoxidil are known to be effective against male-pattern baldness (androgenic alopecia) by male hormones in baldness therapy, but the treatment effect disappears within a few months after discontinuation of treatment. In contrast, finasteride and propecia are therapeutic agents for the treatment of benign prostatic hyperplasia and leukoplakia. Type II 5-reductase (type II 5), an enzyme that converts testosterone to dihydrotestosterone (DHT) -reductase) (Kaufman et al., 1999). This enzyme is present in the hair papilla cells of the hair follicle base, and finasteride and propecia are effective for male hair loss.

On the other hand, hair follicles are remodeled themselves through cycles of anagen, catagen, telogen, and exogen (Paus and Cotsarelis, 1993). During decline, hair follicles undergo programmed cell death, that is, apoptosis, which reduces size and enters the pauses (Cotsarelis, 1997). Hair follicles are regenerated at the beginning of the next growing season, requiring the activation of stem cells in the swollen hair follicles (Cotsarelis et al., 1990). Proliferating stem cells form new hair follicle matrix during the early growth period, and hair shaft and inner root sheath are derived from relatively undifferentiated basal cells (Oshima et al., 2001). The size and length of hair follicle stem are proportional to the size of hair follicle and the duration of growth period, respectively. The cyclic growth of these hair follicles is regulated by various growth factors.

On the other hand, the most common forms of hair loss include male pattern baldness or general baldness, telogen effluvium, cancer-induced hair loss, and alopecia areata. C57BL / 6 C3H / HeJ mice are used as model animals for hair loss studies. This C57BL / 6 mouse is characterized by black body hair and spontaneous alopecia. In the C57BL / 6 C3H / HeJ mouse, melanocytes are present only in the hair follicles, and the synthesis of melanin is in good agreement with the hair growth cycle to determine the hair growth cycle (Pals et al., 1989; Mller-Rver et al., 2001). This C3H / HeJ mouse has histologic and immunohistological characteristics similar to human alopecia areata (Sundberg et al., 1994). Female C3H / HeJ mice over 6 months of age develop alopecia areata with a frequency of 20%, with lymphocytes infiltrating around the hair follicle and effective in steroids, similar to the effect of human immunotherapy (Freyschmidt-Paul et al. , 1999).

As described above, various studies such as providing various causes for hair loss and developing a therapeutic agent or a functional composition for treatment have been actively conducted, but a solution for obtaining excellent effects has not been proposed.

Japan Patent JP2009-203199 (2009.09.10.) Korean Patent No. 10-0594342 (Bulletin of Mar. 3, 2006) Effect of herbal extracts on hair growth in C3H / HeJ mice. Journal of Biomedical Research. 2008.9 (3). pp.57-64

It is an object of the present invention, which is devised to overcome the above-described problems, to provide a method for treating at least one of sea tangle extract, broccoli sprout extract, alfalfa extract, colbai extract, And to provide a composition for improving dandruff removal, hair loss prevention, and hair growth promotion that can prevent hair loss and promote hair growth.

In order to achieve the above object, the composition for improving dandruff removal and hair loss prevention and hair growth promotion according to the present invention is characterized in that the extract of the cystatin, the sea tangle, and the broccoli sprout are mixed in a ratio of 3 (± 0.2): 1 ): 0.5 (+/- 0.07) parts by weight.

In addition, it was also found that a composition obtained by further mixing at least one of 0.1 (± 0.03) parts by weight of various extracts, 0.1 (± 0.03) parts by weight of alfalfa extract 0.1 (± 0.03) parts by weight of collavi extract and 0.1 (± 0.03) parts by weight of resveratrol .

The extracts were mixed with 5.4 mg / kg of various extracts, 5.4 mg / kg of alfalfa extract, 5.4 mg / kg of collavi extract, , And 5.4 mg / kg of resveratrol.

INDUSTRIAL APPLICABILITY As described above, according to the present invention, the sea tangle extract and the broccoli sprout extract are mixed at a certain ratio to the hairless tobacco extract to promote hair growth.

In addition, by further mixing at least one of the various extracts, alfalfa extract, cola extract, and resveratrol, it is possible to maximize hair loss prevention and hair growth promotion by enhancing blood circulation improvement, providing hair nutrients, and dissolving thrombus .

In addition, the composition in which these components are mixed has an excellent effect on suppression of dandruff on the scalp and anti-inflammatory effect.

The composition of the present invention is not only in the form of a shampoo or a hair tonic, but also in a variety of functional foods such as tablets or beverages to prevent hair loss and activate hair growth.

BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate preferred embodiments of the invention and, together with the description, serve to further the understanding of the technical idea of the invention, It should not be interpreted.
The present invention relates to a composition for improving dandruff removal and hair loss prevention and a composition for promoting hair growth, which comprises a cystatin, a seaweed extract, a broccoli sprout extract, a variety of extracts, an alfalfa extract, a collavi extract, A graph showing the growth rate of hair growth for each test substance including the composition according to the ratio of single or mixture, the normal control group and the comparative substance group for 21 days.
Fig. 2 is a photograph showing the degree of hair growth by time of the experimental mouse on the 21st day in Fig.
FIG. 3 is a photograph showing the tissue change of the experimental mouse on day 21 in FIG. 1; FIG.
FIGS. 4A and 4B are graphs showing γ-GTP activity on the 14th day and 21st day in FIG.
FIG. 5 is a graph showing ALP activity measured after application or oral administration of the test substance of FIG. 1 to a mouse. FIG.
FIG. 6 is a graph showing the amount of EGF expression measured after application or oral administration of the test substance of FIG. 1 to a mouse. FIG.
FIG. 7 is a graph showing the amount of VEGF expression measured after application or oral administration of the test substance of FIG. 1 to a mouse. FIG.
FIG. 8 is a photograph showing the change of hair density after oral administration of the test substance of FIG. 1 orally after oral administration of the test substance to the normal hair loss patients.
FIG. 9 is a graph showing changes in hair density after 8 weeks and 16 weeks after oral administration of the test material of FIG.
FIG. 10 is a photograph showing changes in hair thickness after 16 weeks of oral administration of the test substance of FIG.
FIG. 11 is a graph showing changes in hair thickness after 8 weeks and 16 weeks after oral administration of the test material of FIG.
FIG. 12 is a graph showing the visual evaluation scores of dandruff and inflammation and hair loss improvement by the testees after 8 weeks and 16 weeks after the oral administration of the test material of FIG. 1 to the patients who had formalized hair loss.
FIG. 13 is a graph showing the subjective scores of dandruff and inflammation and alopecia improvement by the test subjects after 8 weeks and 16 weeks after the oral administration of the test material of FIG. 1 to the patients who had been prescribed formaldehyde.
FIG. 14 is a photograph showing the results of evaluating the anticancer activity inhibitory effect according to the sole or blending ratio of the cilostazol extract, seaweed extract, and compositions 1 and 2 contained in the composition according to the present invention.
FIG. 15 is a graph showing the results of evaluating inhibitory effect of increased macrophage production of macrophages on RAW 264.7 cell culture system of the extracts of sea tangle, seaweed extract, and compositions 1 and 2 contained in the composition according to the present invention.
FIG. 16 is a graph showing the results of inhibition of nitrogen concentration and inhibition of prostaglandin E ₂ content in inflammation-inducing substances in air pockets, which are included in the composition according to the present invention. Fig.
FIG. 17 is a graph showing the results of inflammatory cell infiltration of infiltration of inflammatory cells into the cytochalasinus tuberosum extract, sea tangle extract, and compositions 1 and 2 contained in the composition according to the present invention (inflammatory cell infiltration inhibition) This is the picture shown.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, the detailed description of known functions and configurations incorporated herein will be omitted when it may unnecessarily obscure the subject matter of the present invention.

The composition for improvement of dandruff removal and hair loss prevention and hair growth promotion according to the present invention comprises Cistanche tubulosa extract, kelp extract and broccoli sprout extract as main ingredients, and various extracts, alfalfa extract, cola extract Non-extract, resveratrol is used as auxiliary material. Each material and mixing ratio will be described in detail below.

First, the sheath tincture extract is mixed with 3 (± 0.2) parts by weight. Such a cystathanum tubuloosa extract is a powder form extracted from a cystathanum tubulosa by extraction with 80% ethanol, sterilization and drying. Here, it is distributed in the ovine grassland of the cystotypes, and it is called "desert ginseng" because it is thick and thick in meat quality, and its section is oily and glossy. It has also been reported that the cytotoxic agent TNF- and IL-4 production, which are the major cytokinases required for the isolation of nitric oxide (NO) from inflammatory cells, has been reported to be lowered, and also has a strong anti-inflammatory effect in animal experiments Yamada et al., 2010; Kyung et al., 2012). In addition, Sysanthus tetuberus exhibited an excellent effect on the improvement of blood circulation by lowering the blood cholesterol level, and the effectiveness of blood circulation promotion, vasodilation, hair growth, dandruff and inflammation improvement I could confirm.

In addition, sea tangle extract (including fucoidan) is mixed at 1 (± 0.1) parts by weight. This kelp extract was obtained by washing, drying and grinding kelp, hot water extraction at 60 ° C for one day, filtering with 0.45 μm membrane filtration, concentration, sterilization, spraying, drying and concentration. Recently, kelp extract has been reported to have excellent thrombolytic efficacy in the prevention and improvement of ischemic diseases (Min et al., 2012a, 2012b). This seaweed extract is a sulfate polysaccharide complex that can be obtained from seaweeds such as Laminaria japonica, Undaria pinnatifida Sporophyll and Cladosiphon okamuranus in many countries. . In previous studies, seaweed extracts reported antioxidant, anticoagulant, anticancer and anti-inflammatory effects (Feldman et al., 1999; Wang et al., 2010). Therefore, the usefulness of seaweed extracts for inflammatory diseases, ischemia, decreased immunity, and tumors has attracted the attention of researchers (Bojakowski et al., 2001; Li et al., 2005). This kelp extract was able to confirm the efficacy of the composition of the present invention for the maintenance and production of healthy hair by enhancing the immunity mechanism of the scalp and hair follicle through blood circulation improvement, anticoagulation and thrombolysis.

In addition, the broccoli bud extract is mixed with 0.5 (0.07) parts by weight. This broccoli sprout extract was obtained by washing, drying and pulverizing broccoli sprouts which are a popular vegetable rich in vitamin C, K, A and fiber, extracting with 70% ethanol at 40 ℃, sterilization, drying and concentrating. The broccoli sprout extract of the present invention contains sulforaphane as a main component. This sulforaphane is more abundant in buds than in broccoli adults (Fahey et al., 2002) and has anti-cancer and anticancer effects through induction of anti-cancer enzymes, apoptosis of cancer cells, inhibition of cell damage and angiogenesis inhibition In the present invention, the composition of the present invention has been reported to have an effect of enhancing and detoxifying the immune system (Fahey et al., 2002; Haristoy et al., 2003; Yanaka et al., 2009) Enhancement of ingredients, improvement of dandruff, dandruff and inflammation were confirmed in the experiments described below.

Then, the colorful extract is mixed with 0.1 (0.03) parts by weight. The various extracts were obtained by extracting, sterilizing, drying and concentrating the variety of green, yellow vegetables belonging to the cruciferous group with 70% ethanol. A variety of vitamins, vitamins or vitamins, and vitamin A is a precursor to the content of carotene is two times the amount of spinach, 100 g of dietary supplement contains 80% of the adult daily vitamins needed to help blood, blood It is known to have functionality to improve circulation. The various extracts obtained from these varieties were found to be effective in the hair health and hair growth through the hematopoietic action and the enhancement of blood circulation promotion in the composition of the present invention.

Further, the alfalfa extract is mixed with 0.1 (0.03) parts by weight. This alfalfa extract was obtained by extracting, sterilizing, drying and concentrating alfalfa, which is a perennial herbaceous plant belonging to the soybean family called alfalfa, in 70% ethanol. Alfalfa has a protein content of more than 25% and has a very good composition of other nutrients (nitrogen, phosphorus, potassium, calcium, boron, sulfur, molybdenum, magnesium, etc.). Recently, a kind of phytoestrogen Alfalfa, which contains a large amount of quercetin and coumesterol, has been increasingly used as a result of its lowering of cholesterol, the improvement of skin beauty and hypertension, and so on. The alfalfa extract obtained from these alfalfa was found to be effective in lowering cholesterol and blood pressure as well as enhancing nutritional content of hair and improving hair growth.

In addition, the cola extract is mixed with 0.1 (0.03) parts by weight. This cola bean extract is a two-year-old vegetable of the genus Papaveraceae, Brassica oleracea (Kohlarbi), which is extracted from 70% ethanol, which is also known as turnip cabbage or caliber cabbage, Lt; / RTI > Cobra is rich in anthocyanins, carotenoids, vitamin C and glucosinolates, which are known to have excellent anticancer effects. They are rich in vitamin C and potassium compared to ordinary vegetables such as cabbage It has been known that it is effective for dieting because it has abundant dietary fiber with low calorie content, and has antimutagenic effect, and has synergistic effect on anti-inflammation and hair growth. .

Then, resveratrol is mixed with 0.1 (0.03) parts by weight. This resveratrol is extracted from grapes. The grapes belong to Rhamnale and Vitaceae, and the fruit is used for reproduction or processed into beverage, wine, jam, grape seed oil, and the like. So far, grape-based studies have shown that they have cardiovascular disease, anti-cancer activity, and immunomodulating activity. Among the physiologically active substances exhibiting such efficacy, resveratrol is a representative substance, and phenolic acid, catechin, flavonoid, proanthocyanidin, and the like are present in various forms. When cultivating grapes, grape leaves are usually pruned, usually discarded. However, young grape leaves can be used not only for edible purposes but also for antioxidative and antiinflammatory effects, thus preventing and improving human diseases. Here, resveratrol was washed with fresh water immediately, and dried in a dryer at 60 to 70 ° C for a day. 100 g of grape leaves were extracted with 1,000 ml of 80% ethanol for one week, and then filtered and washed with a rotary vacuum concentrator And dried at -70 ° C in a freeze dryer (Eyela, FDU-2100, Tokyo Rikakikai Co., Tokyo, Japan). Such resveratrol is excellent in thrombolytic effect and is superior in safety and activity as a metabolite of edible plant compared to conventional thrombolytic agent and can be administered orally unlike conventional thrombolytic agents administered mainly through intravenous injection, , Pulmonary vascular diseases, and the like, and it has been confirmed that it is particularly effective in improving blood circulation and promoting hair growth.

[Animal experiment]

According to the present invention, a mixture of 3: 1: 0.5: 0.1: 0.1: 0.1: 0.1: cysteine body extract, seaweed extract, broccoli sprout extract, various extracts, alfalfa extract, collavi extract and resveratrol The composition is made. Various test materials, including the composition of the present invention, were prepared for the experiment and the resultant contrast of the composition according to the present invention. As a test substance, a normal control group of saline, 3% of minoxidil, and 54 mg / kg of cystatin pertussis extract 1, 18 mg / kg of kelp extract 1, 162 mg / kg 2, and kelp extract 2 of 54 mg / kg were prepared. In addition, the composition of the present invention was divided into composition 1 (low dose) and composition 2 (high dose), and composition 2 was prepared by injecting each material about 3 times as much as composition 2. The composition 1 was prepared in the same manner as in Example 1 except that 54 mg / kg of cystathanum tuberosus, 18 mg / kg of sea tangle extract, 9 mg / kg of broccoli sprout extract, 1.8 mg / kg of various extracts, 1.8 mg / kg of alfalfa extract, 1.8 mg / kg. Composition 2 contained 162 mg / kg of cystathanum tuberosum, 54 mg / kg of seaweed extract, 27 mg / kg of broccoli sprout extract, 5.4 mg / kg of various extracts, 5.4 mg / kg of alfalfa extract, 5.4 mg / kg of collavi extract, 5.4 mg / kg, and stored at about 4 ° C until use.

1. Experimental animals

Experimental animals of the composition according to the present invention were male C57BL / 6 mice at 5 weeks of age and male ICR mice for investigation of hair growth from Orient Bio (Korea) (Horizontal) × 200 (vertical) × 145 (height) (mm) size mouse cage. The environment of the laboratory was adjusted to 23 ± 2 ℃, 55 ± 10% relative humidity, 12 times / hour of ventilation, 12 hours of illumination period, and illumination of 150 ~ 300 Lux. Purina Rat Chow, a pellet-type solid feed for laboratory animals, was supplied and received from Biopia (Korea, Inc.), and the drinking water was allowed to ingest sterile purified water freely.

At 6 weeks of age, the dorsal part of the mouse was removed with an electric razor to induce the dormant hair roots to grow (Hattori and Ogawa, 1983). In other words, if the pink hair was found to be pink after waiting for the epilating skin to uniformly pink, the remaining hair was shaved lightly so that it would not scratch the skin with a razor (Lee et al., 2006; Kim et al., 2009).

2. Test substance

2-1. Normal control group

On the next day of shaving, saline was applied to the shaved area 1-2 times a day for 21 days. The result of the hair growth is shown in Fig.

2-2. Comparative material group

On the next day after shaving, 3% of minoxidil was applied to the depilated skin with a brush about one time per day for about 21 days for 21 days. The result of the hair growth is shown in Fig.

2-3. Systannic To Tuberosa Extract 1

54 mg / kg of Sutentanum tubulosa extract was orally administered once a day for 21 days. The result of the hair growth is shown in Fig.

2-4. Seaweed extract 1

18 mg / kg of kelp extract was orally administered once a day for 21 days. The result of the hair growth is shown in Fig.

2-5. Sutentanus tubulosa extract 2

162 mg / kg of Sutentanus tubulosa extract was orally administered once a day for 21 days. The result of the hair growth is shown in Fig.

2-6. Sea tangle extract 2

54 mg / kg of kelp extract was orally administered once a day for 21 days. The result of the hair growth is shown in Fig.

2-7. Composition 1

Kg of extracts of broccoli sprouts, 1.8 mg / kg of various extracts, 1.8 mg / kg of alfalfa extract, 1.8 mg / kg of collavi extract, 1.8 mg / kg of extracts of resveratrol Was orally administered once a day for 21 days. The result of the hair growth is shown in Fig.

2-8. Composition 2

Kg of alfalfa extract, 5.4 mg / kg of collagen extract, 5.4 mg / kg of resveratrol, 5.4 mg / kg of alfalfa extract, Was orally administered once a day for 21 days. The result of the hair growth is shown in Fig. Here, the capacity of each of the ingredients in Composition 1 and Composition 2 was calculated by applying a weight-to-body surface conversion ratio (Km = 37 / mouse Km = 3) from 10 mg / 70 kg and 300 mg /

3. Analysis of hair growth by age

The back region of the experimental animals was photographed before the administration of the test substance (0 day) and every 3 days after the administration to determine the hair growth effect. At this time, as a camera, a computerized image analysis system (Imageinside (company name), Korea) was used, and the hair growth rate of a regrowth area / shaved area per skin area Respectively.

3-1. Analysis of hair growth by time

Compared with the normal control group, the comparative substance group showed rapid hair growth promoting effect. In detail, on the 21st day, excellent hair growth effect was promoted by 92.4% of 29.2% of the normal control group (see FIGS. 1, 2A and 2B) ).

In addition, seaweed extract 1 had lower hair growth efficacy than the 3% minoxidil comparative substance group, but hatching was promoted in seaweed extract 2, reaching 69% on day 21 (see FIGS. 1, 2C and 2D).

Similar to seaweed extract 2, cystanthus tobacco extract 1 promoted hair growth to 73.6% on day 21 and 80.4% on day 21 of cystatin torrent extract 2 (FIG. 1, 2E and 2F).

The composition 1 was similar to the sea tangle extract 2, and no synergistic effect of the ingredients was observed even on the 21st day. On the other hand, it was confirmed that the synergistic effect of each material was observed in Composition 2, and was similar to the result of 3% of minoxidil in the comparative substance group on the 21st day (see FIGS. 1, 2G and 2H).

4. Histological examination

On the 14th and 21st days after the start of the test, the hairy areas and the skin of the delayed areas were removed and fixed with 10% formalin solution. The tissue was embedded in paraffin, and 5 tissue sections were prepared. Hematoxylin-eosin staining was performed to observe the number and histological changes of the hair follicles by optical microscope.

4-1. Histological examination results

The fidelity of the hair follicle was remarkably improved in the skin histopathological findings on the 14th day and 21st day of the comparative substance group compared to the normal control group (see FIG. 3A) (see FIG. 3B).

The fidelity of these hair roots was lower than that of the comparative substance group, but it was also improved in the seaweed extracts 1 and 2 (see Figs. 3C and 3D), and it was also improved in the suture extracts 1 and 2 and the composition 1. 3E to 3G)

In particular, it was confirmed that the composition 2 was improved more than that of the comparative substance group (Fig. 3H).

5. Measurement of GTP (Glutamyl Transpeptidase) and ALP (Alkaline Phosphatase) Activity

The extracted skin tissue was immediately frozen in liquid nitrogen, and then 4 times a phosphate buffer (pH 7.4) was added and homogenized. The homogenate was centrifuged at approximately 3,000 rpm for approximately 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 PNPP-DEAV ( P- nitrophenolphosphate) (Handiiski et al., 1994; Kim et al., 2009).

5-1. Results of GTP and ALP activity measurements

The GTP activity, which is known to induce hair roots to grow in the growth medium, was increased by 90 ~ 120% as compared with that of the normal control group as in the case of 14 days (FIG. 4A) and 21 days (FIG. 4B). In the case of sea tangle extract 2, the deviation was severe but the GTP activity tended to increase. Especially, in the case of the cystadenum tubulosa extract 2, a significant increase was observed at 14 days. In addition, the compositions 1 and 2 showed a synergistic effect that was prominent on day 21 compared to day 14.

Also, ALP activity, which is known as an animal skeletal growth and angiogenesis index, showed a significant increase in both the 14th day (FIG. 5A) and the 21th day (FIG. 5B) in comparison with the normal control group. In addition, kelp extract 1, 2 and cisternatum tubulosa extract 1, 2 also increased ALP activity in a dose dependent manner. Compounds 1 and 2 were found to be higher than those of seaweed extracts 1 and 2 and cis-tootrobula extracts 1 and 2.

6. EGF (Epidemal Growth Factor) and VEGF (Vascular Endothelial Growth Factor) expression level measurement

EGF and VEGF were measured using an enzyme-linked immunosorbent assay (ELISA) kit (R & D System, Minneapolis, USA) (Cross et al., 2003; Kim et al., 2009). That is, the production of two growth factors was quantitated by sandwich enzyme immunoassay using mouse EGF and VEGF specific polyclonal antibodies according to the manufacturer's guide.

6-1. Results of measurement of EFG and VEGF expression levels

Analysis of EFG promoting growth of hair follicles revealed that compared to the normal control group, the comparative substance group increased 3.5 times over both 14 days (Fig. 6A) and 21 days (Fig. 6B). In addition, seaweed extracts 1 and 2 showed a slight increase on the 14th day but a significant increase on the 21st day. In addition, the ascites of the cystathanum tubuloosa extract 1 was weak, but the cystathanum tubulosa extract 2 exerted an excellent effect on the 14th and 21st days. Here, as shown in the compositions 1 and 2, the synergistic effect between the six materials was confirmed by significantly increasing the EFG content.

On the other hand, when the angiogenic factor VEGF was analyzed, compared with the normal control group, the comparative substance group increased about 80% at 14 days (Fig. 7A) and about 50% at 21 days (Fig. 7B). In addition, seaweed extracts 1 and 2 did not show any effect, and Sytantanthus tubulosa extracts 1 and 2 showed significant VEGF synergistic effect. In the compositions 1 and 2, a synergistic effect was confirmed on the increase of VEFG on the 14th day.

7. Statistical processing

All measurements were expressed as mean standard error. When significance was observed after one-way analysis of variance (ANOVA), the significance between the test substances was tested at the level of P <0.05 through Turkey`s test.

[Clinical Experiment]

1. Test substance

(Hereinafter referred to as "MK"), which is prepared at a ratio of 3: 1: 0.5: 0.1: 0.1: 0.1: 0.1, is added to the extracts of Siltanthus truffilia, Seaweed extract, Broccoli sprout extract, -R7 or HGF-R7), a capsule containing 260 mg of the control maltodextrin was provided to the experimental group to compare the experimental results according to the dose. The control group was given two daily doses of Sisotanthus tuberosum extract, seaweed extract, broccoli sprout extract, colorful extract, alfalfa extract, cola extract, and placebo capsules containing no resveratrol. This is summarized in [Table 1] below.

Test Products Contrast product product name MK-R7 Placebo chief ingredient Systanchus tubulosa extract, seaweed extract, broccoli sprout extract, colorful extract, cola bean extract, resveratrol extract Maltodextrin Product Characteristics capsule capsule usage Twice a day, once a capsule Twice a day, once a capsule

2. Density and diameter of hair (thickness)

Clinical investigators affected by hair loss were shaved 2 mm in length with a 1 cm diameter circular shape near the crown. The density and diameter of the patient's hair were measured before the use of MK-R7 and 16 weeks after the phototrichogram (non-invasive imaging device (Folloscope) 4.0, Lead M) Were used for the measurement. The density of the hair was measured by counting the number of hairs in the 1 cm ² area, and the diameter of the hair was measured by calculating the average value of the measured hair diameters.

2-1. Change in hair density (unit n / cm ² ) Results (primary efficacy endpoint)

The mean of the hair density of the test group was 159.56 ± 28.34 (see FIG. 8A) before consumption of MK-R7 and 182.84 ± 32.96 (see FIG. 8B) after 16 weeks of consumption. The average of the hair mean densities of the control group is 150.18 ± 39.57 (see FIG. 8C) before consumption and 160.53 ± 37.55 (see FIG. 8D) after 16 weeks of consumption. All patients who participated in clinical trials experienced increased hair density. Here, the degree of change of hair density after 16 weeks was increased by 10.35 ± 20.08 in the control group and by 23.29 ± 24.26 in the test group, which is higher in the test group than in the control group. Thus, it can be seen that the increase in the hair density of the test group was statistically significant with the control group and fertilizer (see FIG. 9).

2-2. Results of variation of hair diameter (thickness, unit mm) (secondary efficacy evaluation variable)

The average hair diameter of the test group was 0.063 + 0.014 before consumption of MK-R7 (see FIG. 10A) and 0.086 + 0.018 (FIG. 10B) after 16 weeks. The control group was 0.071 ± 0.018 before consumption of MK-R7 (see Figure 10C) and 0.077 ± 0.015 after 16 weeks (see Figure 10D). Both control and test groups showed increased hair diameter. Both groups showed an increase in hair diameter. However, after 16 weeks, the test group showed a statistically significant increase in hair diameter as compared to the control group (see FIG. 11).

3. Researcher's Score and Subject's Subjective Score

At 8 weeks and 16 weeks, the researchers were provided with photographs of the head side of the test subjects at the top of the standard to check for improvement of hair loss symptoms and dandruff and scalp inflammation. The expert judged the degree of improvement on the 5-point scale (-1: worse, 0: no improvement, +1: fine improvement, +2: improvement, +3: remarkable improvement). In addition, questions were asked at 8 and 16 weeks to identify subjective improvement effects of test subjects on alopecia symptoms and improvement of dandruff and scalp inflammation, and subjects were assessed for improvement on a 5-point scale (-1: dissatisfied , 0: indifference, +1: somewhat satisfied, +2: satisfied, +3: very satisfied).

3-1. The researcher's visual assessment of dandruff, inflammation and hair loss improvement

Assessment by blinded surveys giving visual scores of clinical improvement of hair loss showed improvement in the mean scores of the two groups, with the test group receiving 0.52 ± 0.66 after 8 weeks and the control group receiving 0.24 ± 0.63. Also, after 16 weeks, the test group received 0.64 ± 0.78 and the control group received 0.51 ± 0.82, indicating an increase in the score, but the statistical difference between the two groups was not significant (see FIG. 12A). After 16 weeks, the scores of the researchers in each group showed a statistically significant difference in the scores given in the test and control groups. In addition, after 8 weeks, the test group received a mean score of 0.48 ± 0.62 and the control group received 0.30 ± 0.60 for visual improvement of the improvement effect of dandruff and inflammation. There was no significant statistical difference between the two groups, although there was a slight improvement in both groups. After 16 weeks, the test group received an average score of 0.68 ± 0.64, the control group received 0.35 ± 0.62, and the test group and control group showed significant differences (see FIG. 12B).

3-2. Patient's subjective alopecia improvement satisfaction score

The subjective rating of subjective hair loss improvement satisfaction was 2.55 ± 1.02 in the test group and 2.41 ± 1.04 in the control group after 8 weeks and 2.82 ± 1.01 in the test group and 2.49 ± 1.06 in the control group after 16 weeks , Proving that most participants are satisfied or very satisfied. However, there was no statistically significant difference between the two groups (see FIG. 13A). Regarding subjects' satisfaction with improvement of scalp dandruff and inflammation, the mean score of the test group was 2.21 ± 1.02 and the mean score of the control group was 2.02 ± 1.07 after 8 weeks. In addition, after 16 weeks, the mean score of the test group was 2.65 ± 1.04 and the mean score of the control group was 2.13 ± 1.05. The mean scores given to the test group and the control group thus showed a statistically significant difference (see FIG. 13B).

[Dandruff and inflammation related]

As an immune response to external antigens, macrophages have been implicated in inflammatory responses such as tumor necrosis factor-alpha, TNF-alpha, interleukin-1 beta, IL-1 beta and interleukin 6 Secrete the cytokines. Such cytokines induce inflammatory cells such as granulocytes, mononuclear leukocytes, lymphocytes, and mast cells around the injured tissue so that these cells can remove the antigen, . However, excessive infiltration and activation of inflammatory cells exacerbate tissue damage, leading to edema (vascular leakage) and pain, which are the main symptoms of the inflammatory reaction.

Tumor necrosis factor receptors are important factors in inducing nitric oxide synthase (iNOS) genes in some cells. Activation of nitric oxide synthase promotes the production of nitrogen (NO), which causes inflammation, as well as dilating blood vessels, killing bacteria. On the other hand, the tissue damage causes the production of arachidonic acid by phospholipases in cell membrane phospholipids. This arachidonic acid is activated by cyclooxygenase (COXs) prostaglandins, PGs). Coke produced in -Ⅱ (cox-Ⅱ) excessive prostaglandin E ₂ (prostaglandin E ₂₎ by activation induces talkie ness between various proinflammatory. Nitric oxide (TNF-α-NO) and Cox-II-prostaglandin E2 (COX-Ⅱ-PGE2) pathways are the major pathogenetic pathways of NGF and NGF as nitric oxide and prostaglandin E ₂ are important factors in inflammation and pain induction. These two pathways are inhibited by corticosteroids and non-steroidal anti-inflammatory drugs (NSAIDs), respectively. Although there are a variety of steroid drugs and NSAIDs for the treatment of inflammatory diseases, these drugs have side effects in the immune system, gastrointestinal tract, kidney, liver, central nervous system, blood pressure, and cardiovascular system. Therefore, there is a need to minimize these side effects by replacing these chemical agents or by administering them in combination with natural products.

1. Dustpanic culture and killing efficacy

For the experiment of the composition according to the present invention, M. furfur ATCC 15421 was cultivated in Sabouraud dextrose agar (SDA) after being distributed from American Type Cluture Collection (ATCC; Manassas, USA). As an agar-dilution assay, each test substance was continuously diluted from 2 mg / mL to 0.004 mg / mL in a Sabouraud medium to prepare a medium. The cells were plated on paraffin oil and inoculated at 30 μL in accordance with 1.5 × 10 ⁸ cells (MacFaland Nl. 0.5). After culturing for 72 hours, the colonies were observed to determine whether the cells were grown Respectively.

2. Cell culture and determination of nitrogen

RAW 264.7 macrophages were purchased from ATCC and contained 10% fetal bovine serum (Sigma, St. Louis, USA) and 100 U / mL of antibiotics (Sigma) And cultured in Dulbecco's modified Eagle's medium (DMEN; Sigma) in a humidified 5% CO ₂ incubator at 37 ° C. In order to evaluate the effect of kelp extract and cis-tootrobula extract on nitrogen (NO) production, RAW 264.7 cells (1 × 10 ⁶ cells / mL) were treated with 1-320 μL / mL of the test substance for 5 minutes 10 U / mL of interferon-y (interferon-y, IFN-y) and 10 μg / mL of lipopolysaccharide (LPS) were added and cultured for 24 hours. It was measured using the nitrite (nitrite, NO ₂₎ oxidation reaction of nitrogen as an index of the nitrogen, the grease reagent (griess reagent) the same amount of culture solution of 1% sulfanyl amide (sulfanilamide), 0.1% in 2.5% phosphoric acid (phosphoric acid) The mixture was mixed with naphthylethylenediamide and allowed to react at room temperature for 10 minutes. Nitrogen content was measured at 540 nm and quantified using a standard quantitative curve of sodium nitrite (NaNo ₂ ).

3. Experimental animals

Seven week old male ICR mice were received from Korea BioLink (company name, voice, Korea) and used for about 1 week after purification process. Animals were housed in cages (220 x 200 x 145 mm), four in each. The environment of the animal laboratory was controlled at 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 illumination of 150-300 Lux. Purine Rat Chow, a pellet type solid feed for laboratory animals, was supplied and received from Biopia (company name, Gunpo, Korea), and the drinking water was allowed to drink sterilized purified water freely.

4. Administration of test substance and induction of inflammation

(18mg / kg) or sea tangle extract (54mg / kg), Sisotanthus bulurosa extract 1 (54mg / kg) or Sisotecum tubulosa extract 2 (162mg / kg) kg), mixture 1 (18 + 54 mg / kg) or mixture 2 (54 + 162 mg / kg) of a mixture of sea tangle extract and cystathanum tubulosa extract and composition 1 or composition 2 described above was orally administered daily for 7 days . The low dose of kelp extract 1 and cisternatum tubulosa 1 were calculated by applying body weight to body surface conversion ratio (adult Km = 37, mouse km = 3) from 100 mg / 70 kg and 300 mg / kg, respectively. In addition, the high dose of kelp extract 2 and cystathanum tubulosa 2 were three times the volume of seaweed extract 1 and cystatin tetrofosloxy 2, both of which were administered alone or in combination, and the maximum dose was set at 5 times. The highest synergistic effect among the test substances was evaluated. An air-pouch was made by injecting 10 mL of sterile air subcutaneously under the back of the mouse at the start of the experiment. After 2 and 5 days, 5 mL of air was further infused to keep the bladder stable. One hour after the last test substance administration, 1 mL of carrageenan (1% in saline; Sigma) or only the solvent And injected into the pocket.

5. Exudative analysis

After 6 hours of injection of carrageenan, the bladder was rinsed with 1 mL of cold saline and the net volume of the lavage fluid was recorded. The number of inflammatory cells such as neutrophils, mononuclear leukocytes, and lymphocytes was measured with a coulter counter. The inflammatory mediators nitrogen and prostaglandin E ₂ were measured by enzyme immunoassay (EIA) using a grease reagent (Sigma) and a correlate-EIA kit (Assay Eesigns, Ann Arbor, Ann ArboUSA).

6. Histopathological examination

The skin and subcutaneous tissues including the inner tissue of the bladder were collected and fixed with 10% neutral formalin solution. The tissues were embedded in paraffin, slides were prepared, stained with hematoxyling-eosin, and inflammatory findings were observed under an optical microscope.

7. Statistical processing

All measurements were expressed as mean ± standard error. When significance was observed after one-way analysis of variance (ANOVA), the significance between groups was tested at the level of P <0.05 through Turkey`s test.

8. Results

As a result of evaluating the inhibitory effect of Staphylococcus aureus extracts 1, 2, kelp extract 1, 2 and compositions 1, 2 on Streptococcus mutans furfur proliferation, . By showing the effect of inhibiting the proliferation of. furfur, the dicumer killing efficacy was confirmed (see Fig. 14).

In addition, the extracts 1 and 2 and the sea tangle extracts 1 and 2 did not affect the nitrogen liberation from the increased macrophages. In contrast, the compositions 1 and 2 were found to have anti-inflammatory effects by significantly inhibiting nitrogen production from a concentration of 32 μg / mL (see FIG. 15).

As a result of measurement of the nitrogen content in the air bladder for the cystatin tufrugosa extract 1, 2, kelp extract 1, 2, and composition 1, 2, the kelp extract 1, 2 and the cystatin torso extract 1, While Compositions 1 and 2 lowered the nitrogen concentration to an excellent level and further decreased dose-dependently. In a similar manner, the prostaglandin E ₂ content increased by the inflammatory agent (carrageenan) was also synergistically inhibited by Compositions 1 and 2 (see FIG. 16).

As a result of evaluating the inhibitory effect on the inflammatory cell infiltration of the Sutentanthus tuboulos extract 1, 2, the kelp extract 1, 2 and the compositions 1, 2, , And the anti-inflammatory effect was further enhanced by the administration of the compositions 1 and 2 (see FIG. 17).

In conclusion, the antimicrobial efficacy against M. furfur, which is a representative dandruff strain, was evaluated in order to examine the effect of anti-inflammatory effect on skin damage and itching by dandruff formation. , And compositions 1 and 2 showed a complete proliferation inhibitory effect at a concentration of 1 mg / mL or more. In addition, the inhibitory effects of Sibutan tuberosum extract and kelp extract on inflammatory cell inhibitory effect were slight, whereas compositions 1 and 2 significantly inhibited macrophage activation. This has been demonstrated in the suppression of nitrogen, a major inflammatory reaction secreted from macrophages, in cell culture systems and animal experiments. In addition, Prostaglandin E ₂ , which is an important inflammatory mediator, also showed an excellent inhibitory effect on the cystanthin tobacco extract and seaweed extract, and the composition 1, 2 further enhanced the anti-inflammatory effect. As described above, it has been confirmed that compositions 1 and 2 can improve symptoms such as dandruff formation and inflammation when applied in oral form and application form.

It will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. It is therefore to be understood that the above-described embodiments are to be considered in all respects as illustrative and not restrictive. The scope of the present invention is defined by the appended claims, rather than the detailed description, and all changes or modifications derived from the meaning and scope of the claims and equivalents of the claims are to be construed as being included within the scope of the present invention do.

Claims

(± 0.2): 1 (± 0.1): 0.5 (± 0.07): 0.1 (± 0.03) of the extracts of Shisotan tufurosa, seaweed extract, broccoli sprout, colorful extract, alfalfa extract, columbia extract and resveratrol, A functional food composition for improving dandruff removal and prevention of hair loss and promoting hair growth by mixing 0.1 (± 0.03): 0.1 (± 0.03): 0.1 (± 0.03)

delete

(± 0.2): 1 (± 0.1): 0.5 (± 0.07): 0.1 (± 0.03) of the extracts of Shisotan tufurosa, seaweed extract, broccoli sprout, colorful extract, alfalfa extract, columbia extract and resveratrol, Wherein the composition is prepared by mixing 0.1% (± 0.03): 0.1 (± 0.03): 0.1 (± 0.03) by weight.