Composition for promoting hair growth and application thereof

Abstract

The present invention provides a composition for promoting hair growth and uses thereof, the composition comprising: a mixture comprising a Polygonum multiflorum extract and nano-silver particles, and a copper-blue peptide, the weight / volume ratio of the copper-blue peptide to the composition for promoting hair growth being 0.1 mg / ml to 0.2 mg / ml, the weight / volume ratio of the copper-blue peptide to the composition for promoting hair growth being 0.1 mg / ml to 0.2 mg / ml, and the weight / volume ratio of the copper-blue peptide to the composition for promoting hair growth being 0.1 mg / ml to 0.2 mg / ml. The mixture containing the fleece-flower root extract and the nano-silver particles comprises the nano-silver particles, the fleece-flower root extract, a stabilizer and a diluent. The invention also provides an application of the mixture for preparing a medical composition for promoting hair growth. The composition for promoting hair growth can effectively promote hair growth and promote generation of new hair follicles.

Description

Technical Field

[0001] The 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 the use of the composition in promoting hair growth. Background Art

[0002] Alopecia areata is a skin disease in which hair is lost from the scalp or other parts of the body. It usually begins as small, smooth, round bald patches on the scalp and progresses to alopecia totalis or alopecia univeralis.

[0003] It is generally believed that alopecia areata is caused by the body's immune system attacking the hair follicles. Other observations have pointed out that it may be caused by the naturally produced hormone dihydroxytestosterone (DHT) in the body that shortens the hair growth period, making the grown hair too fine and difficult to see, leading to hereditary male pattern baldness. If it continues to develop, it will result in a receding hairline in men.

[0004] Alopecia areata can occur in all people, regardless of gender or race. It can start in adolescence. Epidemiologically, alopecia affects 1.7% of the world's population, including more than 4.7 million people in the United States alone. The above situation may have an impact on a person's appearance, self-confidence, quality of life, and work or school.

[0005] According to U.S. Patent Publication 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 effects is still the goal of the current technical field of the present invention to develop and study. Summary of the invention

[0006] In order to solve the above technical problems, the object of the present invention is to provide a composition for promoting hair growth, which comprises: a mixture comprising a Polygonum multiflorum extract and nano-silver particles and a blue copper peptide, wherein the weight / volume ratio of the blue copper peptide to the composition for promoting hair growth is 0.1 mg / mL (mg / mL) to 0.2 mg / mL, and the mixture comprising a Polygonum multiflorum extract and nano-silver particles comprises nano-silver particles, a Polygonum multiflorum extract, a stabilizer and a diluent.

[0007] Although it is currently known that nanosilver particles and Polygonum multiflorum extract can be used to improve alopecia areata, the present invention can bring unexpectedly better and more excellent hair growth promoting effect by further combining blue copper peptide and controlling the ratio of the mixture including Polygonum multiflorum extract and nanosilver particles to blue copper peptide.

[0008] According to the present invention, the blue copper peptide is a complex structure with copper as the central atom surrounded by molecules of glycine-histidine-lysine triamino acids. In one embodiment, the blue copper peptide of CAS No. 49557-75-7 is used. In another embodiment, the blue copper peptide of CAS No. 89030-95-5 is used.

[0009] 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. 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.

[0010] Preferably, the particle size of the nanosilver particles is less than or equal to 50 nanometers (nm), and more preferably, the particle size of the nanosilver particles is less than 20nm. Even more preferably, the particle size of the nanosilver particles is less than 10nm. Preferably, the particle size of the nanosilver particles is greater than or equal to 1nm. According to the present invention, the particle size is a diameter.

[0011] Preferably, 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. Preferably, the weight / volume ratio of the Polygonum multiflorum extract to the mixture comprising the Polygonum multiflorum extract and the nano-silver 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 nano-silver particles is 0.91 mg / mL to 1.82 mg / mL.

[0012] 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.

[0013] Preferably, the Polygonum multiflorum extract is a Polygonum multiflorum root extract.

[0014] Preferably, the preparation method of the root extract of Polygonum multiflorum is: (a) mixing Polygonum multiflorum 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.

[0015] Preferably, the Polygonum multiflorum is dried Polygonum multiflorum.

[0016] 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%.

[0017] Preferably, the solid-liquid ratio (w:v) of the Polygonum multiflorum to the alcohol solvent is 1:1 to 1:30. For example, when the solid-liquid ratio (w:v) of the Polygonum multiflorum to the alcohol solvent is 1:1, it means that 100 grams (g) of the Polygonum multiflorum is mixed with 100 mL of the alcohol solvent; when the solid-liquid ratio (w:v) of the Polygonum multiflorum to the alcohol solvent is 1:30, 100 g of the Polygonum multiflorum is mixed with 3000 mL of the alcohol solvent.

[0018] According to the present invention, the Polygonum multiflorum extract used in the embodiment is purchased from GREENTECHBIOTECHNOLOGIES, and the extraction method is to grind 100g of dried Polygonum multiflorum root and soak it in 1000mL of 50 volume percent ethanol aqueous solution, extract for 48 hours, filter and separate to obtain a Polygonum multiflorum root extract, and then reduce the pressure to concentrate the Polygonum multiflorum root extract, freeze-dry to obtain a Polygonum multiflorum root extract weighing 10g. The extraction rate of the Polygonum multiflorum root extract is 10%.

[0019] Preferably, the stabilizer is a protein, a peptide, sodium borohydride or polyvinylpyrrolidone. Among them, the protein can be human serum protein or transferrin. Preferably, the stabilizer is polyvinylpyrrolidone (PVP), which can effectively stabilize the nanosilver particles. Preferably, based on the total volume of the composition for hair growth, the concentration of the PVP is 0.1 millimoles per liter (mM) to 1 mM. 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.

[0020] To achieve the aforementioned object, the present invention further provides a use of a composition for promoting hair growth for preparing a pharmaceutical composition for promoting hair growth, wherein the composition for promoting hair growth is as described above.

[0021] Preferably, the subject to which the pharmaceutical composition is administered is a mammal, such as a human, a dog, a cat or a rodent.

[0022] Preferably, the pharmaceutical composition is applied to the skin surface by applying.

[0023] Preferably, the pharmaceutical composition is administered to a human once to three times a day for at least 7 weeks.

[0024] The hair growth promoting composition of the present invention can effectively promote the hair growth of nude mice, has a high hair coverage rate and the effect of promoting the proliferation of new hair follicles. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 The average hair coverage of mice applied with Example 1 and Comparative Examples 1 to 4 from the 1st to the 7th week;

[0026] FIG. 2A to FIG. 2H The photos are taken under an optical microscope of the skin hair follicles of mice smeared with Example 1 and Comparative Examples 1 to 4 at 7 weeks. The scale bar in the figure is 100 micrometers. DETAILED DESCRIPTION

[0027] In order to have a clearer understanding of the technical features, purposes 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 construed as limiting the applicable scope of the present invention.

[0028] The present invention will be further described by the following examples, which are not intended to limit the present invention. Those skilled in the art may make some improvements and modifications without departing from the scope of the present invention.

[0029] Reagent Description:

[0030] Nanosilver particle aqueous solution, product model: DA04-21, CAS No. 7440-22-4, purchased from Liwei Nano Technology Co., Ltd., nanosilver particle concentration is 0.00085wt% to 0.0015wt%, water is 99.9985wt% to 99.99915wt%;

[0031] Polyvinylpyrrolidone, product model: SI-PVP40-500G, CAS No.9003-39-8, purchased from Youhe Trading Co., Ltd.;

[0032] 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%;

[0033] Blue copper peptide, product model: 1310CSB00010050, CAS No.49557-75-7, was purchased from Abike Co., Ltd., Taiwan Province, China.

[0034] Preparation Example 1 Preparation of a mixture containing Polygonum multiflorum extract and nanosilver particles

[0035] 100 milliliters (mL) of a nanosilver aqueous solution (DA04-21, Liwei Nanotech Co., Ltd., with a concentration of nanosilver particles of 0.00085 wt% to 0.0015 wt%) were poured into a beaker with a handle and stirred with a magnet, wherein the nanosilver particles in the nanosilver aqueous solution had a particle size of less than 10 nm. 2.5 grams (g) of polyvinyl pyrrolidone were slowly added to the nanosilver aqueous solution in batches, mixed and stirred until the addition was complete, and then continued to stir until it was completely dissolved and the mixture was clear. In addition, a Polygonum multiflorum extract (GREENTECH BIOTECHNOLOGIES, 305558, with a concentration of Polygonum multiflorum extract 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 Polygonum multiflorum extract and nanosilver particles. Among them, 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.

[0036] Example 1 Composition for promoting hair growth

[0037] The mixture containing the Polygonum multiflorum extract and the nano-silver particles is mixed with the blue copper peptide aqueous solution with a concentration of 0.5 mg / mL, and the volume ratio of the two is 2.5:1 to obtain the hair growth promoting composition of this embodiment. In this embodiment, 110 mL of the mixture containing the Polygonum multiflorum extract and the nano-silver particles and 44 mL of the blue copper peptide aqueous solution are selected, wherein the concentration of the blue copper peptide in the hair growth promoting composition is 0.14 mg / mL.

[0038] Comparative Examples 1 to 4 Compositions for Promoting Hair Growth

[0039] Comparative Examples 1 to 4 are similar to Example 1, except that the volume ratio of the mixture containing Polygonum multiflorum extract and nanosilver particles to the blue copper peptide aqueous solution with a concentration of 0.5 mg / mL is used. The volume ratios of the Polygonum multiflorum extract and the nanosilver mixture to the blue copper peptide aqueous solution in Comparative Examples 1 to 4 are 20:1, 10:1, 5:1, and 1:1, respectively. The composition and ratio of each comparative example and example are shown in Table 1 below.

[0040] Table 1 Composition of Example 1 and Comparative Examples 1 to 4

[0041]

[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 randomly divided into groups after adaptive feeding for one to two weeks, with 5 mice in each group. A silicone splint (Grace Bio-Labs, Bend, OR, USA, product model: SKU: 476687) was sewn on the back of each mouse, with an outer diameter, inner diameter and thickness of 24 mm, 20 mm and 0.5 mm, respectively. The hair growth promoting composition of different embodiments, comparative examples or control groups was administered to the mouse skin in the silicone splint on the back of the mouse every day, and the volume was up to 40 microliters (μL) for a total of 7 weeks. In this test example, the mixture containing Polygonum multiflorum extract and nanosilver particles (without blue copper peptide) of Preparation Example 1 was administered to the back of the mouse as the positive control group; the negative control group was not administered any drug to the back of the mouse as the negative control group; and the blue copper peptide control group was administered to the back of the mouse as a 0.5 mg / mL blue copper peptide aqueous solution. The mice were weighed once a week to confirm their health status. The hair growth patterns of mice were photographed on days 0, 7, 14, 21, 28, 35, and 42, that is, weeks 1, 2, 3, 4, 5, 6, and 7 after the start of the experiment. The mice were anesthetized with 2.5% isoflurane and placed under a stereomicroscope photography system (SZN71 Microscope system, Sunny, Hangzhou, China). The built-in photography software was used to take the photos, and then the Image J software was used to analyze the hair coverage. The range of the inner diameter of the silicone splint attached to the back of the mouse (i.e., the range of drug administration) was selected and the total administration area was calculated. Then, the hair growth range was selected again and the hair growth range area was calculated. The hair coverage formula is (hair growth range area ÷ total administration area) × 100%. The hair coverage results of Example 1, Comparative Examples 1 to 4 and the control group are shown in the figure. Figure 1 shown.

[0044] according to Figure 1The average results of the hair coverage of all mice in each group show that there was no obvious hair coverage in the dosing range of each group from the 1st to the 3rd week. At the 7th week, compared with the hair coverage of Comparative Examples 1 to 4 and the control group of 30.6%, 19.8%, 16.4%, 11.8% and 22.0%, respectively, the hair coverage of Example 1 was 61.8%, which was the best result. The hair coverage results of each group at the 7th week were further analyzed by two-way ANOVA (Tukey test) to determine whether there was a significant difference compared with the positive control group. The results are shown in Table 2 below. According to the results of Table 2, at the 7th week, only the hair coverage results of Example 1 were significantly different from those of Comparative Examples 1 to 4 and the control group.

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

[0046] Group Significant difference Comparative Example 1 No significant difference Comparative Example 2 No significant difference Comparative Example 3 No significant difference Example 1 *Significantly different Comparative Example 4 No significant difference

[0047] Test Example 2 Calculation of hair follicle count

[0048] After all mice in Test Example 1 were sacrificed, the mouse skin tissues were embedded in paraffin and cut into 3 μm thick sections on glass slides for histopathological analysis by hematoxylin and eosin stain (H&E) and immunohistochemistry (IHC). Hematoxylin and eosin stain (H&E stain) was performed, and the stained slides were scanned using a slide panoramic scanner (OLYMPUS / VS200). Under the condition of 80× magnification, 2 fields of view were randomly selected for each section for hair follicle observation and calculation. The results are shown in the figure. FIG. 2A to FIG. 2H .

[0049] FIG. 2A to FIG. 2H They are slices of negative control group, positive control group, blue copper peptide control group, comparative example 1, comparative example 2, comparative example 3, embodiment 1, and comparative example 4, respectively, and the arrows on each group of figures are examples of circular hair follicles. Figure 2G As shown, the number of hair follicles in the slices of the group treated by Example 1 was significantly better than that of the other groups ( FIG. 2A to FIG. 2F , Figure 2H ), and the distribution of hair follicles is more dense. Therefore, the composition for promoting hair growth of the present invention can effectively promote the proliferation of new hair follicles.

[0050] In summary, the hair growth promoting composition of the present invention can effectively promote the hair growth of nude mice, has a high hair coverage rate and increases the generation of new hair follicles.

[0051] It will be apparent to those skilled in the art that various modifications and variations may be made in accordance with the present invention without departing from the scope and spirit of the present invention. Although the present invention has described certain preferred embodiments, it should be understood that the present invention should not be unduly limited to such specific embodiments. In fact, various modifications that are obvious to those skilled in the art in the described modes of carrying out the present invention are also included within the scope of the claims of the present invention.

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, wherein 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, wherein: 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, wherein: 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, wherein: The Polygonum multiflorum extract is a Polygonum multiflorum root extract.

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

7. The composition for promoting hair growth according to claim 1, wherein: 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, wherein: 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, wherein The subject to which the pharmaceutical composition is administered is a mammal.

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

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