Composition For Preventing Hair Loss Or Promoting Hair Regrowth

Abstract

The present disclosure relates to a composition for preventing alopecia and accelerating hair growth, including an ingredient inducing new development of hair or accelerating hair growth. The composition for preventing alopecia and accelerating hair growth reduces a period of transition from a telogen stage to an anagen stage in the hair growth cycle to accelerate hair growth and to delay a transition to a catagen stage, and thus provides excellent effects of preventing alopecia and accelerating hair growth. In addition, the composition is safe to the human body, causes no side effect and provides excellent effects of accelerating proliferation of dermal papilla cells and enhancing hair elasticity.

Claims

1. A composition for preventing alopecia or accelerating hair growth, comprising escin as an active ingredient.

2. The composition for preventing alopecia or accelerating hair growth according to claim 1, wherein the content of the active ingredient is 0.001-10 wt %, based on the total weight of the composition.

3. A composition for preventing alopecia or accelerating hair growth, comprising at least one compound selected from rhaponticin, rhein, chrysophanol and physicon-8-O-d-glucopyranoside, or a salt, hydrate or solvate thereof, as an active ingredient.

4. The composition for preventing alopecia or accelerating hair growth according to claim 3, which comprises at least two of the compounds.

5. The composition for preventing alopecia or accelerating hair growth according to claim 3, wherein the compound comprises: (i) chrysophanol; and (ii) at least one compound selected from rhein, rhaponticin and physicon-8-O-d-glucopyranoside.

6. The composition for preventing alopecia or accelerating hair growth according to claim 3, wherein the compound comprises rhaponticin, rhein, chrysophanol and physicon-8-O-d-glucopyranoside.

7. The composition for preventing alopecia or accelerating hair growth according to claim 3, wherein the content of the active ingredient is 0.00001-50 wt %, based on the total weight of the composition.

8. The composition for preventing alopecia or accelerating hair growth according to claim 1, which accelerates proliferation of dermal papilla cells.

9. The composition for preventing alopecia or accelerating hair growth according to claim 1, which enhances hair elasticity.

10. The composition for preventing alopecia or accelerating hair growth according to claim 1, which is a pharmaceutical composition.

11. The composition for preventing alopecia or accelerating hair growth according to claim 1, which is a non-medical composition.

12. The composition for preventing alopecia or accelerating hair growth according to claim 1, which is a cosmetic composition.

13. A hair or scalp care product comprising the composition for preventing alopecia or accelerating hair growth as defined in claim 1.

14. The hair or scalp care product according to claim 13, which is selected from the group consisting of a hair growth treatment, scalp clinic agent, scalp scaling agent, scalp massaging agent, scalp care agent, cleaner, shampoo, tonic, hair conditioner, hair lotion, gel, pack, cream, essence, powder, spray, oil, soap, ointment, hair styling agent, hair dye and hair perm agent.

15. A method for preventing alopecia or accelerating hair growth in a scalp of a subject, comprising a step of administering to the subject the composition according to claim 1.

16. A method for preventing alopecia or accelerating hair growth in a scalp of a subject, comprising a step of administering to the subject the composition according to claim 3.

Description

DESCRIPTION OF DRAWINGS

[0051] FIG. 1 illustrates the results of analysis about the effect of amplifying activity of Wnt/β-catenin promoter of escin in dermal papilla cells.

[0052] FIG. 2 illustrates the results of the test for determining an increase in expression of β-catenin caused by escin in dermal papilla cells.

BEST MODE

[0053] Hereinafter, the present disclosure will be explained in more detail with reference to exemplary embodiments. This disclosure may, however, be embodied in many different forms and should not be construed as limited to the exemplary embodiments set forth therein. It will be apparent that these exemplary embodiments are provided so that the present disclosure will be complete and understood easily by those skilled in the art.

PREPARATION EXAMPLE 1

Composition 1 for Treating Alopecia/Accelerating Hair Growth (Hair Tonic)

[0054] Hair tonics including escin according to Examples 1-6 and Comparative Example 1 were prepared in the conventional manner by using the formulations as shown in the following Table 1.

TABLE-US-00001 TABLE 1 Weight ratio (%) Comp. Ingredients Ex. 1 Ex. 1 Ex. 2 Ex. 3 Ex. 4 Ex. 5 Ex. 6 Ethanol 55 55 55 55 55 55 55 Castor Oil 5 5 5 5 5 5 5 Glycerin 3 3 3 3 3 3 3 Escin — 0.01% 0.1% 0.5% 1% 2% 5% Fragrance q.s. q.s. q.s. q.s. q.s. q.s. q.s. and Colorant Purified Balance (Total 100) Water

PREPARATION EXAMPLE 2

Composition 2 for Treating Alopecia/Accelerating Hair Growth (Hair Lotion)

[0055] Hair lotion including escin according to Example 7 was prepared in the conventional manner by using the formulation as shown in the following Table 2.

TABLE-US-00002 TABLE 2 Ingredients Weight ratio (%) Cetostearyl Alcohol 2 Stearyltriethylammonium Chloride 2 Hydroxyethyl Cellulose 0.5 Escin 2 Fragrance and Colorant q.s. Purified Water Balance (Total 100)

EXPERIMENTAL EXAMPLE 1

Effect of Controlling Signals (Wnt/β-catenin) Activated in Dermal Papilla Cells

[0056] In general, Wnt/β-catenin signals activated in dermal papilla cells are generated while a transition from a catagen stage to an anagen stage occurs in the hair growth cycle, i.e. during a period from beginning of hair growth to an activation stage of hair growth. In addition, in a telogen stage and catagen stage, Wnt/β-catenin signals become weak or disappear to cause degradation of hair follicles and depilation. In this Experimental Example, it is determined how escin contributes to amplification of Wnt/β-catenin signals.

[0057] As a positive control which amplifies Wnt/β-catenin signals, wnt3a protein was used. In addition, dimethyl sulfoxide (DMSO) was used as a negative control. In a 96-well culture plate, approximately 3×10.sup.4 of Wnt reporter HEK293SA cells were seeded to each well and treatment with escin was carried out at a concentration of 5 μg/mL, 10 μg/mL and 20 μg/mL. Then, a luciferase assay kit (E1960) available from Promega Corp. was used to carry out reporter analysis according to the test method provided by the company. In addition, luminometer Victor (Perkin Elmer, Waltham, Mass., USA) was used to determine Wnt/β-catenin promoter activity. The analysis results are shown in FIG. 1.

[0058] As can be seen from the test results, escin amplifies Wnt/β-catenin signals by approximately 11 times as compared to the non-treated negative control. The test group including escin shows no concentration-dependent behavior but shows a repetitive clear effect at a concentration of about 20 μg/mL. Particularly, it is shown that escin provides a higher effect as compared to the positive control, wnt3a protein, bound directly to the receptor. Therefore, it can be seen that escin significantly accelerates the activity of the Wnt/β-catenin signal (hair growth-accelerating signal) transmission system, and thus provides an excellent effect of accelerating hair growth.

EXPERIMENTAL EXAMPLE 2

Effect of Accelerating Proliferation of Dermal Papilla Cells

[0059] Human-derived dermal papilla cells (DPCs) were purchased from PromoCell GmbH. DPCs were cultured under the conditions of 37° C. and 5% CO.sub.2 by using a follicle dermal papilla cell growth medium and supplement mix recommended by PromoCell GmbH. The cultured DPCs were pipetted to a 96-well plate at 3,000 cells/well and cultured for 24 hours in the presence of 0.1% of blood serum. Then, the cultured cells were treated with escin at a concentration of 0.2, 0.5, 1.2 and 5 μg/mL for one day and then incubated, and DMSO (vehicle) diluted to 1:1000 in serum-free DMEM was used as a control. After incubation, a cell counting kit (CCK) was used to evaluate proliferation of cells. The culture medium was treated with CCK-8 at a ratio of 1:10, followed by incubation for 1 hour. After 1 hour, the absorbance of each well was determined at 450 nm. All tests were repeated three times and the average of absorbance value was calculated. The results are shown by the percentage of a test group with the result of the control taken as 100. After the test, treatment with escin shows an effect of DPC proliferation, suggesting an excellent effect of proliferation of dermal papilla cells.

TABLE-US-00003 TABLE 3 Treatment Dermal Papilla Cell Test Group Concentration Proliferation Ability (%) Non-treated group — 100.0 Escin 0.2 μg/mL 168.8 0.5 μg/mL 198.7   1 μg/mL 194.4   2 μg/mL 204.8   5 μg/mL 208.3

EXPERIMENTAL EXAMPLE 3

Increase in Expression of β-catenin in Dermal Papilla Cells

[0060] In dermal papilla cells in a catagen stage and a telogen stage, during which Wnt signals are inhibited, β-catenin is decomposed by GSK3-beta. However, in dermal papilla cells in an anagen stage, during which Wnt signals are amplified, activity of GSK3-beta is inhibited and β-catenin is not decomposed, resulting in an increase in expression of β-catenin. Human dermal papilla cells were pipetted to six well plates at about 500,000 cells/well, and treated with escin for 1 day at a concentration of 0.2, 0.5, 1.2 and 5 μg/mL, followed by incubation. After incubation, M-PER lysis buffer was used to cause lysis of the cells and the same amount of cell lysate was loaded on SDS-PAGE gel. The cell lysate loaded on SDS-PAGE gel was transferred to a nitrocellulose membrane and anti-β-catenin antibody was attached to the nitrocellulose membrane to detect protein. GAPDH was used as a control for correcting protein content. The amount of β-catenin increased by escin was determined. The results are shown in FIG. 2.

[0061] After the test, it can be seen that the group treated with escin shows a significant increase in amount of β-catenin, which suggests that Wnt/β-catenin signals are increased in dermal papilla cells.

EXPERIMENTAL EXAMPLE 4

Determination of Hair Growth Effect of Composition 1 (Hair Tonic) for Treating Alopecia/Accelerating Hair Growth

[0062] Composition 1 (hair tonic) for treating alopecia/accelerating hair growth according to the present disclosure was determined in terms of hair growth effect for 105 male and female subjects having a significantly smaller number of hair strands as compared to a normal state or suffering from alopecia and having weak hair. The 105 males and females were divided into 7 groups each including 15 subjects and treated with Comparative Example 1 and Examples 1-6. Each sample was used on hair and scalp five times per week for 6 months. After using each sample, improvement or deterioration was evaluated with a five-grade scoring system in terms of hair thickness, degree of crowding, elasticity and overall evaluation (improvement: +5, +4, +3, +2, +1, no improvement: 0, deterioration: −1, −2, −3, −4, −5). The evaluation was based on the average of the results of examination by interview after using each sample for 6 months. The results are shown in the following Table 4.

TABLE-US-00004 TABLE 4 Hair Degree of Overall Sample Thickness Crowding Elasticity Evaluation Comp. Ex. 1 −0.5 ± 0.2   0.1 ± 0.2 −0.1 ± 0.2   −0.21 ± 0.2    Ex. 1 2.1 ± 0.2 1.5 ± 0.2 2.5 ± 0.2 2.0 ± 0.2 Ex. 2 3.2 ± 0.1 2.6 ± 0.2 2.6 ± 0.5 2.8 ± 0.2 Ex. 3 4.5 ± 0.3 4.3 ± 0.2 3.8 ± 0.3 4.2 ± 0.1 Ex. 4 4.6 ± 0.2 4.3 ± 0.1 4.8 ± 0.3 4.6 ± 0.2 Ex. 5 4.8 ± 0.2 4.6 ± 0.1 4.3 ± 0.2 4.6 ± 0.1 Ex. 6 3.1 ± 0.4 2.1 ± 0.3 2.3 ± 0.6 2.5 ± 0.2

[0063] As can be seen from the above results, Examples 1-6 including escin shows a high effect of preventing alopecia and accelerating hair growth. This demonstrates that the composition for preventing alopecia and accelerating hair growth including escin as an active ingredient according to the present disclosure is significantly effective for treating alopecia.

EXPERIMENTAL EXAMPLE 5

Determination of Hair Growth Effect of Composition 1 (Hair Tonic) for Treating Alopecia/Accelerating Hair Growth

[0064] Composition 1 (hair tonic) for treating alopecia/accelerating hair growth according to the present disclosure was determined in terms of hair growth effect for 105 male and female subjects having a significantly smaller number of hair strands as compared to a normal state or suffering from alopecia and having weak hair. The 105 males and females were divided into 7 groups each including 15 subjects and treated with Comparative Example 1 and Examples 1-6. Each sample was used on hair and scalp five times per week for 6 months. After using each sample, researcher evaluation through clinical photographs was carried out for Comparative Example and Examples, 2, 4 and 6 months after applying each sample, with a 3-grade evaluation system of ‘good’, ‘slight’ and ‘no change’ (good: 50-70% improved, slight: 20-50% improved, no change). Evaluation of the number of hair strands per unit area and average hair thickness through phototrichogram was carried out for Comparative Example and Examples, 4 and 6 months after applying each composition. The results are shown in the following Table 5.

TABLE-US-00005 TABLE 5 Phototrichogram Researcher Evaluation through Clinical Photographs Number of Hair Average Hair No change Slight Good Strands per Unit Thickness (person) (person) (person) Area (No./cm.sup.2) (μm) Sample 2M 4M 6M 2M 4M 6M 2M 4M 6M 0M 4M 6M 0M 4M 6M Comp. 13 13 12 2 1 2 0 0 1 372 ± 42 375 ± 53 379 ± 24 60 ± 5 60 ± 3 62 ± 2 Ex. 1 Ex. 1 9 8 8 5 6 4 1 1 3 370 ± 27 382 ± 37 397 ± 38 65 ± 4 65 ± 5 67 ± 3 Ex. 2 8 7 7 4 6 5 3 2 3 327 ± 20 367 ± 39 384 ± 26 60 ± 4 65 ± 8 69 ± 3 Ex. 3 6 6 4 3 2 3 6 7 8 356 ± 28 364 ± 19 398 ± 22 64 ± 6 69 ± 3 76 ± 7 Ex. 4 4 4 2 8 6 3 3 5 10 334 ± 33 383 ± 33 403 ± 54 61 ± 8 64 ± 3 72 ± 2 Ex. 5 3 3 3 8 5 1 4 7 11 322 ± 24 387 ± 46 397 ± 28 64 ± 7 68 ± 5 75 ± 5 Ex. 6 6 6 5 3 2 3 6 7 7 384 ± 41 394 ± 18 408 ± 47 62 ± 6 62 ± 4 69 ± 5

[0065] As can be seen from the above results, Examples 1-6 including escin shows a high effect of preventing alopecia and accelerating hair growth. This demonstrates that the composition for preventing alopecia and accelerating hair growth including escin as an active ingredient according to the present disclosure is significantly effective for treating alopecia.

[0066] The formulation example, such as hair tonic or hair lotion, is merely an exemplary embodiment of the composition for accelerating hair growth according to the present disclosure, and it is apparent to those skilled in the art that the scope of the composition according to the present disclosure is not limited to the above formulations.

PREPARATION EXAMPLE 3

Composition 3 for Treating Alopecia (Hair Tonic)

[0067] At least one of rhaponticin, rhein, chrysophanol and physicon-8-O-glucopyranoside was used to prepare hair tonic in the conventional manner according to the composition of the following Table 6.

TABLE-US-00006 TABLE 6 Weight ratio (%) Comp. Comp. Ingredients Ex. 2 Ex. 3 Ex. 8 Ex. 9 Ex. 10 Ex. 11 Ex. 12 Ethanol 55 55 55 55 55 55 55 Castor Oil 5 5 5 5 5 5 5 Glycerin 3 3 3 3 3 3 3 Active 0 Minoxidil Rhaponcitin Rhein Chrysophanol Physicon- Rhaponcitin, Ingredient 2 μg/mL 10 μg/mL 10 μg/mL 10 μg/mL 8-O-d- Rhein, glucopyranoside Chrysophanol, 10 μg/mL and Physicon- 8-O-d- glucopyranoside Each 2.5 μg/mL Fragrance q.s. q.s. q.s. q.s. q.s. q.s. q.s. and Pigment Purified Balance (total 100) Water

PREPARATION EXAMPLE 4

Composition 4 for Treating Alopecia (Hair Lotion)

[0068] At least one of rhaponticin, rhein, chrysophanol and physicon-8-O-glucopyranoside was used to prepare hair lotion in the conventional manner according to the composition of the following Table 7.

TABLE-US-00007 TABLE 7 Ingredients Weight ratio (%) Cetostearyl alcohol 2 Stearytriethylammonium chloride 2 Hydroxyethyl cellulose 0.5 Rhaponticin, Rhein, Chrysophanol or 0.001 Physicon-8-O-glucopyranoside Fragrance and Pigment q.s. Purified Water Balance (total 100)

EXPERIMENTAL EXAMPLE 6

Effect of Accelerating Proliferation of Dermal Papilla Cells

[0069] Human-derived dermal papilla cells (DPCs) were purchased from PromoCell GmbH. The DPCs were cultured in a DMEM medium (Hyclone Inc., Utah, USA) containing 5% of fetal bovine serum (FBS; Gibco, NY, USA), 10 units/mL of penicillin and 100 μg/mL of streptomycin at 37° C. under 5% of CO.sub.2. The cultured DPCs were pipetted to a 96-well plate at 3,000 cells/well and cultured under the condition of 0.1% of serum for 24 hours. Then, the cultured cells were treated with 10 μg/mL of each of the samples of rhaponticin, rhein, chrysophanol, physicon-8-O-glucopyranoside and combinations thereof as shown in the following Table 8 for one day and then incubated, and DMSO (vehicle) diluted to 1:1000 in serum-free DMEM was used as a control and 2 μg/mL of minoxidil was used as a positive control . After incubation, a cell counting kit (CCK) was used to evaluate proliferation of cells. The culture medium was treated with CCK-8 at a ratio of 1:10, followed by incubation for 1 hour. After 1 hour, the absorbance of each well was determined at 450 nm. All tests were repeated three times and the average of absorbance value was calculated. The results are shown by the percentage of a test group as the result of the control is taken as 100.

[0070] After the test, treatment with rhaponticin, rhein, chrysophanol, physicon-8-O-glucopyranoside or a combination thereof shows an excellent effect of accelerating proliferation of dermal papilla cells (Table 8).

TABLE-US-00008 TABLE 8 Dermal Papilla Treatment Cell Proliferation Test Group Concentration Ability (%) Non-treated group — 100.0 Minoxidil 2 μg/mL 187.2 Rhaponticin 10 μg/mL 162.7 Rhein 10 μg/mL 152.7 Chrysophanol 10 μg/mL 152.0 Physicon-8-O-d-glucopyranoside 10 μg/mL 148.3 Rhaponticin and Chrysophanol each 5 μg/mL 172.2 Rhein and Chrysophanol each 5 μg/mL 168.5 Chrysophanol and Physicon-8-O- each 5 μg/mL 160.5 d-glucopyranoside Rhaponticin, Rhein, Chrysophanol each 2.5 μg/mL 189.4 and Physicon-8-O-d-glucopyranoside

EXPERIMENTAL EXAMPLE 7

Effect of Controlling Dermal Papilla Cell Activity Signals

[0071] In general, it is known that VEGF, IGF-1, FGF, or the like, secreted in dermal papilla cells induce an anagen stage of hair. In addition, versican is known as a marker of an anagen stage of hair and DKK-1 is known as a marker of a catagen stage of hair. Therefore, effects of rhaponticin, rhein, chrysophanol and physicon-O-d-glucopyranoside upon the signals of accelerating an anagen stage and those of inducing a catagen stage in dermal papilla cells were examined according to the present disclosure.

[0072] In a 6-well culture plate, dermal papilla cells were seeded to each well at 1×10.sup.5 and treated with 10 μg/mL of each of the samples of rhaponticin, rhein, chrysophanol, physicon-O-d-glucopyranoside and combinations thereof as shown in the following Table 10. Then, real-time PCR was used to determine expression of each of VEGF, versican and DKK-1 by using Taqman® probe (Thermo Fisher, Massachusetts, USA) as shown in the following Table 9. The results are shown in the following Table 10.

TABLE-US-00009 TABLE 9 Genes Taqman ® Probe Analysis ID VEGF Hs00900055_m1 Versican Hs00171642_m1 DKK-1 Hs00183740_m1

[0073] As a result, it can be seen that rhaponticin, rhein, chrysophanol and physicon-O-d-glucopyranoside increase expression of VEGF, which is a growth factor inducing an anagen stage of hair, and versican as a marker of an anagen stage, and reduce expression of DKK-1, which is a factor inducing a catagen stage of hair (Table 10).

TABLE-US-00010 TABLE 10 Expression of Genes (%) Test Group/Treatment Concentration VEGF Versican DKK-1 Non-treated group 100 100 100 Minoxidil 2 μg/mL 110 134 128 Rhaponticin 10 μg/mL 210 130 112 Rhein 10 μg/mL 144 154 76 Chrysophanol 10 μg/mL 250 158 123 Physicon-8-O-d-glucopyranoside 10 μg/mL 164 175 75 Rhaponticin 5 μg/mL and 235 150 111 Chrysophanol 5 μg/mL Rhein 5 μg/mL and Chrysophanol 5 μg/mL 250 158 65 Chrysophanol 5 μg/mL and Physicon-8-O- 264 192 109 d-glucopyranoside 5 μg/mL Rhaponticin 2.5 μg/mL, Rhein 2.5 μg/mL, 295 188 61 Chrysophanol 2.5 μg/mL and Physicon-8- O-d-glucopyranoside 2.5 μg/mL

[0074] It is shown that rhaponticin, rhein, chrysophanol and physicon-O-d-glucopyranoside have excellent effects of inducing hair growth and inhibiting alopecia. The composition for preventing alopecia and accelerating hair growth according to the present disclosure was prepared with various formulations. Particular formulation examples are described herein. However, the formulation examples described herein are for illustrative purposes only, and it is apparent to those skilled in the art that the scope of the present disclosure is not limited to those formulation examples.

EXPERIMENTAL EXAMPLE 8

Test for Determining Effect of Hair Growth of Composition 3 for Treating Alopecia (Hair Tonic)

[0075] Composition 3 (hair tonic) for treating alopecia obtained from Preparation Example 3 was used for 40 male and female subjects having a significantly smaller number of hair strands as compared to a normal state or suffering from alopecia and having weak hair. The 40 males and females were divided into 4 groups each including 10 subjects and treated with Comparative Examples 2 and 3 and Examples 8-12. Each sample was used on hair and scalp five times per week for 6 months. After using each sample, improvement was evaluated in terms of hair thickness, degree of crowding, elasticity and overall evaluation (significantly improved: +3, improved: +2, slightly improved: +1, no change: 0, slightly deteriorated: −1, deteriorated: −2, significantly deteriorated: −3). The evaluation was based on the average of the results of examination by interview after using each sample for 6 months. The results are shown in the following Table 11.

TABLE-US-00011 TABLE 11 Thickness Degree of Overall Sample of Hair Crowding Elasticity Evaluation Comp. Ex. 2 −0.5 ± 0.2   0.1 ± 0.2 −0.1 ± 0.2   −0.2.1 ± 0.2    Comp. Ex. 3 2.1 ± 0.2 2.3 ± 0.1 2.1 ± 0.3 2.1 ± 0.2 Ex. 8 2.8 ± 0.3 2.7 ± 0.1 2.5 ± 0.4 2.8 ± 0.1 Ex. 9 2.6 ± 0.1 2.8 ± 0.2 2.4 ± 0.3 2.7 ± 0.2 Ex. 10 1.9 ± 0.1 2.6 ± 0.3 2.0 ± 0.2 2.3 ± 0.1 Ex. 11 2.6 ± 0.3 2.1 ± 0.1 2.4 ± 0.1 2.6 ± 0.1 Ex. 12 2.7 ± 0.3 2.9 ± 0.3 2.7 ± 0.3 2.8 ± 0.3

[0076] As can be seen from the above results, Examples using rhaponticin, rhein, chrysophanol and physicon-O-d-glucopyranoside show high effects of preventing alopecia and accelerating hair growth. Thus, it can be seen that the composition according to the present disclosure is significantly effective for treating alopecia.