Hair growth-promoting composition and use thereof

Abstract

Provided is a hair growth-promoting composition, which comprises: a hair growth-promoting composition, which comprises a mixture comprising a Polygonum multiflorum extract and a silver nanoparticle, and a blue copper peptide, wherein the weight / volume ratio of the blue copper peptide to the hair growth-promoting composition is 0.1 mg / mL to 0.2 mg / mL, and the mixture comprising the Polygonum multiflorum extract and the silver nanoparticle comprises the silver nanoparticle, the Polygonum multiflorum extract, a stabilizer, and a diluent. Also provided is use of the hair growth-promoting composition in the preparation of a pharmaceutical composition for promoting hair growth. The hair growth-promoting composition can effectively promote hair growth and enhance new hair follicle generation.

Description

Composition for promoting hair growth and use thereof Technical Field

[0001] The present invention relates to a composition for promoting hair growth, in particular to a composition containing silver particles and a Polygonum multiflorum extract, and also relates to use of the composition in promoting hair growth. Background Art

[0002] Alopecia areata is a skin disorder that causes hair loss on the scalp or other parts of the body. It often begins as small, smooth, round patches of baldness on the scalp and progresses to complete baldness (alopecia totalis) or alopecia univeralis.

[0003] Alopecia areata is generally believed to be caused by the immune system attacking hair follicles. Other observations suggest that the naturally occurring hormone dihydroxytestosterone (DHT) may shorten the anagen phase, making the hair that grows too fine and difficult to see, leading to male pattern baldness. If this condition continues, it can result in a receding hairline.

[0004] Alopecia areata affects everyone, regardless of gender or race. It can begin in adolescence. Epidemiologically, it affects 1.7% of the world's population, including over 4.7 million people in the United States alone. This condition can impact a person's appearance, self-confidence, quality of life, and work or school performance.

[0005] According to U.S. Patent No. 9,295,694, it is currently known that a combination of nanosilver particles and Polygonum multiflorum extract can be used to improve alopecia areata. However, the pursuit of more excellent results is still the goal of the current development and research in the field of the present invention.

[0006] Summary of the Invention

[0007] To solve the above technical problems, the present invention provides a hair growth promoting composition comprising: a mixture comprising a Polygonum multiflorum extract and nanosilver particles; and a copper peptide, wherein the weight / volume ratio of the copper peptide to the hair growth promoting composition is 0.1 mg / mL to 0.2 mg / mL. The mixture comprising a Polygonum multiflorum extract and nanosilver particles comprises nanosilver particles, a Polygonum multiflorum extract, a stabilizer, and a diluent.

[0008] Although nanosilver particles and Polygonum multiflorum extract are currently known to be used to improve alopecia areata, the present invention, by further combining them with copper peptides and controlling the ratio of the mixture containing Polygonum multiflorum extract and nanosilver particles to the copper peptides, can achieve unexpectedly better hair growth-promoting effects.

[0009] According to the present invention, the blue copper peptide is a complex structure with copper as the central atom surrounded by molecules of the amino acids glycine, histidine, and lysine. In one embodiment, the blue copper peptide with CAS No. 49557-75-7 is used. In another embodiment, the blue copper peptide with CAS No. 89030-95-5 is used.

[0010] Preferably, the weight / volume ratio of the nanosilver particles to the mixture comprising the Polygonum multiflorum extract and the nanosilver particles is 0.001 mg / mL to 0.1 mg / mL. More preferably, the weight / volume ratio of the nanosilver particles to the mixture comprising the Polygonum multiflorum extract and the nanosilver particles is 0.005 mg / mL to 0.02 mg / mL. Even more preferably, the weight / volume ratio of the nanosilver particles to the mixture comprising the Polygonum multiflorum extract and the nanosilver particles is 0.0077 mg / mL to 0.014 mg / mL.

[0011] Preferably, the nanosilver particles have a particle size of 50 nanometers (nm) or less. More preferably, the nanosilver particles have a particle size of less than 20 nm. Even more preferably, the nanosilver particles have a particle size of less than 10 nm. Preferably, the nanosilver particles have a particle size of 1 nm or greater. According to the present invention, the particle size is a diameter.

[0012] Preferably, the weight / volume ratio of the Polygonum multiflorum extract to the mixture comprising the Polygonum multiflorum extract and the nanosilver particles is 0.1 mg / mL to 10 mg / mL. Preferably, the weight / volume ratio of the Polygonum multiflorum extract to the mixture comprising the Polygonum multiflorum extract and the nanosilver particles is 0.5 mg / mL to 5 mg / mL. More preferably, the weight / volume ratio of the Polygonum multiflorum extract to the mixture comprising the Polygonum multiflorum extract and the nanosilver particles is 0.91 mg / mL to 1.82 mg / mL.

[0013] Preferably, the weight / volume ratio of the blue copper peptide to the hair growth promoting composition is 0.1 mg / mL to 0.18 mg / mL. Preferably, the weight / volume ratio of the blue copper peptide to the hair growth promoting composition is 0.1 mg / mL to 0.16 mg / mL.

[0014] Preferably, the Polygonum multiflorum extract is a Polygonum multiflorum root extract.

[0015] Preferably, the preparation method of the Polygonum multiflorum root extract is: (a) mixing a Polygonum multiflorum root with an alcohol solvent, extracting at a temperature of 40°C to 60°C for 24 to 72 hours, separating to remove the extracted Polygonum multiflorum, and obtaining a Polygonum multiflorum extract; (b) removing the solvent from the Polygonum multiflorum extract and drying to obtain a Polygonum multiflorum extract.

[0016] Preferably, the Polygonum multiflorum is dried Polygonum multiflorum.

[0017] Preferably, the alcohol solvent may be methanol, ethanol or propanol. The concentration of the alcohol solvent may be 30 volume percent (vt%) to 95 vt%.

[0018] Preferably, the solid-liquid ratio (w:v) of Polygonum multiflorum to the alcohol solvent is 1:1 to 1:30. For example, when the solid-liquid ratio (w:v) of Polygonum multiflorum to the alcohol solvent is 1:1, it means that 100 grams (g) of Polygonum multiflorum is mixed with 100 mL of alcohol solvent; when the solid-liquid ratio (w:v) of Polygonum multiflorum to the alcohol solvent is 1:30, it means that 100 g of Polygonum multiflorum is mixed with 3000 mL of alcohol solvent.

[0019] According to the present invention, the Polygonum multiflorum root extract used in the examples was purchased from GREENTECH BIOTECHNOLOGIES. The extraction method involved crushing 100 g of dried Polygonum multiflorum root and soaking it in 1000 mL of 50% by volume ethanol. After extraction for 48 hours, the extract was filtered and separated to obtain a Polygonum multiflorum root extract. The Polygonum multiflorum root extract was then concentrated under reduced pressure and freeze-dried to obtain 10 g of a Polygonum multiflorum root extract. The extraction yield of the Polygonum multiflorum root extract was 10%.

[0020] Preferably, the stabilizer is a protein, a peptide, sodium borohydride or polyvinyl pyrrolidone. The protein may be human serum albumin or transferrin. Preferably, the stabilizer is polyvinyl pyrrolidone (PVP), which can effectively stabilize the nanosilver particles. Preferably, the concentration of the PVP is 0.1 millimoles per liter (mM) to 1 mM based on the total volume of the hair growth composition. In one embodiment, the concentration of the PVP is 0.57 mM. Preferably, the diluent is selected from the group consisting of water, ethanol, dimethyl sulfoxide, acetonitrile, Hepes buffer, phosphate buffer, Tris buffer, citrate buffer, serum or any other type of physiologically relevant solvent or solution and a combination thereof.

[0021] To achieve the aforementioned objectives, the present invention further provides a composition for promoting hair growth for use in preparing a pharmaceutical composition for promoting hair growth, wherein the composition for promoting hair growth is as described above.

[0022] Preferably, the subject to which the pharmaceutical composition is administered is a mammal, such as a human, a dog, a cat or a rodent.

[0023] Preferably, the pharmaceutical composition is applied to the skin surface by applying it on the skin.

[0024] Preferably, the pharmaceutical composition is administered to a human once to three times a day for at least 7 weeks.

[0025] The hair growth promoting composition of the present invention can effectively promote hair growth in nude mice, has a high hair coverage rate and increases the proliferation of new hair follicles. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] FIG1 shows the average hair coverage of mice treated with Example 1 and Comparative Examples 1 to 4 from week 1 to week 7;

[0027] 2A to 2H are photos of the skin hair follicles of mice smeared with Example 1 and Comparative Examples 1 to 4 at week 7 under an optical microscope. The scale bar in the figures is 100 μm. DETAILED DESCRIPTION

[0028] In order to have a clearer understanding of the technical features, objectives and beneficial effects of the present invention, the technical solution of the present invention is now described in detail below, but it should not be understood as limiting the scope of implementation of the present invention.

[0029] The present invention will be further illustrated by the following examples, which are not intended to limit the present invention. Those skilled in the art may make slight improvements and modifications without departing from the scope of the present invention.

[0030] Reagent Description:

[0031] Nanosilver particle aqueous solution, product model: DA04-21, CAS No. 7440-22-4, purchased from Liwei Nano Technology Co., Ltd., with a nanosilver particle concentration of 0.00085 wt% to 0.0015 wt% and water concentration of 99.9985 wt% to 99.99915 wt%;

[0032] Polyvinylpyrrolidone, product model: SI-PVP40-500G, CAS No. 9003-39-8, purchased from Youhe Trading Co., Ltd.

[0033] Polygonum multiflorum extract, product model: 300558, purchased from GREENTECH BIOTECHNOLOGIES, composition ratio: water: 49.0wt% to 49.5wt%, propylene glycol: 49.0wt% to 49.5wt%, Polygonum multiflorum root extract: 1.0wt% to 2.0wt%;

[0034] Blue copper peptide, product model: 1310CSB00010050, CAS No. 49557-75-7, was purchased from Taiwan Abbic Co., Ltd.

[0035] Preparation Example 1 Preparation of a mixture containing Polygonum multiflorum extract and nanosilver particles

[0036] 100 milliliters (mL) of a nanosilver aqueous solution (Liwei Nanotechnology Co., Ltd., DA04-21, with a nanosilver particle concentration of 0.00085 wt% to 0.0015 wt%) was poured into a beaker with a handle and stirred with a magnet. The nanosilver particles in the nanosilver aqueous solution had a particle size of less than 10 nm. 2.5 grams (g) of polyvinyl pyrrolidone was slowly added to the nanosilver aqueous solution in batches, mixed and stirred until the addition was complete, and then stirred until the polyvinyl pyrrolidone was completely dissolved and the mixture became clear. Separately, a Polygonum multiflorum extract (GREENTECH BIOTECHNOLOGIES, 305558, with a Polygonum multiflorum extract concentration of 1 wt% to 2 wt%) was filtered through a 0.45 micron (μm) filter cup, and 10 mL of the filtered Polygonum multiflorum extract was added to the aforementioned mixture to obtain a mixture comprising the Polygonum multiflorum extract and the nanosilver particles. The weight / volume ratio of the nanosilver particles to the mixture comprising the Polygonum multiflorum extract and the nanosilver particles is 0.0077 mg / mL to 0.014 mg / mL; the weight / volume ratio of the Polygonum multiflorum extract to the mixture comprising the Polygonum multiflorum extract and the nanosilver particles is 0.91 mg / mL to 1.82 mg / mL; and the concentration of polyvinyl pyrrolidone is approximately 0.57 mM.

[0037] Example 1 Composition for promoting hair growth

[0038] The hair growth promoting composition of this example was prepared by mixing a mixture of Polygonum multiflorum extract and nanosilver particles with a 0.5 mg / mL aqueous solution of copper peptide at a volume ratio of 2.5:1. In this example, 110 mL of the mixture of Polygonum multiflorum extract and nanosilver particles and 44 mL of the aqueous solution of copper peptide were used. The concentration of copper peptide in the hair growth promoting composition was 0.14 mg / mL.

[0039] Comparative Examples 1 to 4 Compositions for Promoting Hair Growth

[0040] Comparative Examples 1 to 4 are similar to Example 1, differing only in the volume ratio of the mixture containing Polygonum multiflorum extract and nanosilver particles to the 0.5 mg / mL aqueous copper peptide solution. The volume ratios of the Polygonum multiflorum extract / nanosilver mixture to the aqueous copper peptide solution in Comparative Examples 1 to 4 were 20:1, 10:1, 5:1, and 1:1, respectively. The compositions and proportions of each comparative example and example are shown in Table 1 below.

[0041] Table 1 Composition of Example 1 and Comparative Examples 1 to 4

[0042] Test Example 1 Hair Coverage Experiment

[0043] The following experiment was conducted using Example 1 and Comparative Examples 1 to 4 as samples: Six-week-old nude mice (Nu / Nu mice) were acclimated to feeding for one to two weeks and then randomly divided into groups of five. A silicone splint (Grace Bio-Labs, Bend, OR, USA, SKU: 476687) with an outer diameter, inner diameter, and thickness of 24 mm, 20 mm, and 0.5 mm, respectively, was sewn onto the back of each mouse. A hair growth-promoting composition from each Example, Comparative Example, or control group was administered daily to the skin within the silicone splint for seven weeks. In this test example, the mixture containing the Polygonum multiflorum extract and nanosilver particles (without the copper peptide) from Preparation Example 2 was administered to the back of the mice as a positive control group; no drug was applied to the back of the mice as a negative control group; and a 0.5 mg / mL copper peptide aqueous solution was applied to the back of the mice as a copper peptide control group. The mice were weighed weekly to confirm their health. Hair growth patterns of the mice were photographed on days 0, 7, 14, 21, 28, 35, and 42 (i.e., weeks 1, 2, 3, 4, 5, 6, and 7) after the start of the experiment. Mice were anesthetized with 2.5% isoflurane and placed under a stereomicroscope (SZN71 Microscope system, Sunny, Hangzhou, China). Images were taken using the built-in photography software. Hair coverage was then analyzed using Image J software. The inner diameter of the silicone splint attached to the mouse's back (i.e., the area where the drug was administered) was selected and the total area of ​​drug administration was calculated. The hair growth area was then selected and the area of ​​hair growth was calculated. The formula for hair coverage is (hair growth area ÷ total area of ​​drug administration) × 100%. The hair coverage results for Example 1, Comparative Examples 1 to 4, and the control group are shown in Figure 1.

[0044] The average hair coverage of all mice in each group in Figure 1 shows that no significant hair coverage was observed within the dosing range of any group from Weeks 1 to 3. At Week 7, the hair coverage of Example 1 was 61.8%, the best result, compared to the hair coverage of Comparative Examples 1 to 4 and the control group, which were 30.6%, 19.8%, 16.4%, 11.8%, and 22.0%, respectively. A two-way ANOVA (Tukey test) was used to analyze the hair coverage of each group at Week 7 to determine whether there were significant differences compared to the positive control group. The results are shown in Table 2 below. Table 2 shows that only the hair coverage of Example 1 showed significant differences compared to Comparative Examples 1 to 4 and the control group at Week 7.

[0045] Table 2: Whether there are significant differences between Example 1 and Comparative Examples 1 to 4 compared with the positive control group

[0046] Test Example 2: Calculation of Hair Follicle Number

[0047] All mice in Test Example 1 were sacrificed, and their skin tissues were embedded in paraffin and cut into 3 μm-thick sections on glass slides for histopathological analysis using hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC). H&E staining was performed, and the stained slides were scanned using an OLYMPUS / VS200 panoramic slide scanner. At 80X magnification, two randomly selected fields of view from each section were used for hair follicle observation and enumeration. The results are shown in Figures 2A to 2H.

[0048] Figures 2A through 2H show sections from the negative control group, positive control group, copper peptide control group, Comparative Examples 1, 2, 3, Example 1, and 4, respectively. Arrows in each group indicate circular hair follicles. As shown in Figure 2G , the number of hair follicles in the sections from the group treated with Example 1 was significantly higher than that in the other groups (Figures 2A through 2F, and 2H), and the follicles were more densely distributed. Therefore, the hair growth-promoting composition of the present invention effectively promotes the proliferation of new hair follicles.

[0049] In summary, the hair growth promoting composition of the present invention can effectively promote hair growth in nude mice, has a high hair coverage rate and increases the generation of new hair follicles.

[0050] Various modifications and variations of the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the present invention has been described with certain preferred embodiments, it should be understood that the invention should not be unduly limited to such specific embodiments. In fact, various modifications obvious to those skilled in the art in carrying out the described modes of the invention are intended to be encompassed by the scope of the claims.

Claims

1. A composition for promoting hair growth, comprising: A mixture containing Polygonum multiflorum extract and nano-silver particles and a blue copper peptide. in, The weight / volume ratio of the blue copper peptide to the hair growth promoting composition is 0.1 mg / ml to 0.2 mg / ml, The mixture containing the Polygonum multiflorum extract and nano silver particles comprises nano silver particles, a Polygonum multiflorum extract, a stabilizer and a diluent.

2. The composition for promoting hair growth according to claim 1, in, The weight / volume ratio of the nano-silver particles to the mixture comprising the Polygonum multiflorum extract and the nano-silver particles is 0.001 mg / ml to 0.1 mg / ml.

3. The composition for promoting hair growth according to claim 1, in, The particle size of the nano silver particles is greater than or equal to 1 nanometer and less than or equal to 50 nanometers.

4. The composition for promoting hair growth according to claim 1, in, The weight / volume ratio of the Polygonum multiflorum extract to the mixture comprising the Polygonum multiflorum extract and the nano-silver particles is 0.1 mg / ml to 10 mg / ml.

5. The composition for promoting hair growth according to claim 1, in, The Polygonum multiflorum extract is a Polygonum multiflorum root extract.

6. The composition for promoting hair growth according to claim 1, in, The stabilizer is protein, peptide, sodium borohydride or polyvinyl pyrrolidone.

7. The composition for promoting hair growth according to claim 1, in, The diluent is selected from the group consisting of water, ethanol, dimethyl sulfoxide, acetonitrile, Hepes buffer, phosphate buffer, Tris buffer, citrate buffer, serum and combinations thereof.

8. Use of a composition for promoting hair growth for preparing a pharmaceutical composition for promoting hair growth, in, The hair growth promoting composition is the hair growth promoting composition according to any one of claims 1 to 7.

9. The use according to claim 8, in, The subject to which the pharmaceutical composition is administered is a mammal.

10. The use according to claim 9, in, The pharmaceutical composition is applied to the skin surface by applying.

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