Ginsenoside composition, daily chemical product, and use thereof

Abstract

The present invention belongs to the technical field of daily chemicals and discloses a ginsenoside composition that acts on hair. The composition is any one of ginsenoside compositions N1, N2, N4, N5, and N6, or a combination of multiple ginsenoside compositions. The composition can achieve optimal hair loss prevention and hair growth effects at a relatively low dose, ultimately resulting in a significant reduction in hair loss or thicker hair for the user. At the same time, the present invention also provides uses of the composition and daily chemical products using the composition.

Description

[Technical Field]

[0001] The present invention relates to the technical field of daily chemicals, specifically to ginsenoside compositions, daily chemical products, and their uses. [Background technology]

[0002] Carrot extract is already well known as an active ingredient for preventing hair loss and promoting hair growth, and Japanese hair growth quasi-drugs specify carrot extract as an active ingredient with hair growth properties. Related patent documents are as follows: CN115154387A relates to a method for manufacturing a hair care essence, specifically a hair care essence for promoting hair growth, and its manufacturing method. It has a mild formulation, comprehensive efficacy, and maximizes the retention of the active ingredients and biological activity of plant extracts and multiple proteins. The formulation is designed to enhance hair quality and hair follicle nutrition, improve hair follicle skin differentiation, and enhance hair care and hair loss prevention effects, overcoming the shortcomings of the prior art. The ingredients include glucosylglycerol, dihydroquercetin glucoside, methylpropanediol, hydrolyzed soy protein, hydrolyzed wheat protein, panthenol, tocopheryl acetate, adenosine, carrot extract, peony root extract, larch extract, sodium hyaluronate, and deionized water. The mass percentages of each ingredient are as follows: glucosylglycerol 6-8.5%, dihydroquercetin glucoside 4-9.5%, methylpropanediol 4-8%, hydrolyzed soy protein 0.5-3%, hydrolyzed wheat protein 0.5-3%, panthenol 0.2-1%, tocopheryl acetate 0.2-1%, adenosine 0.2-1%, carrot extract 0.3-1%, and peony root extract 0.3-1%, respectively.

[0003] CN113577003A relates to a three-in-one hair growth shampoo containing core ingredients and auxiliary ingredients. The core ingredients include black sesame, Panax notoginseng, rose, snakeroot, cumin, cherry, Astragalus root, jasmine, ginger, Polygonum multiflorum, ginseng, Angelica sinensis, Dioscorea chinensis pod, black rice, Euonymus gracilis, Poria cocos, and Szechuan ginseng. It also discloses a method for producing a three-in-one hair growth shampoo. The beneficial effects achieved by this invention are that it activates hair follicles, supplies nutrients necessary for hair growth, improves dryness, yellowing, and split ends, and effectively removes dandruff and oiliness. These multiple functions are combined without mutual interference, allowing for consistent use.

[0004] CN113197911A provides a use of a ginsenoside composition in the manufacture of a medicament for preventing and treating hair loss by acting on hair follicular tissue. The ginsenoside composition is composed of ginsenoside Rg1, ginsenoside Re, and ginsenoside CK, of which ginsenoside Rg1 and ginsenoside Re are the main active ingredients, and ginsenoside CK is an auxiliary ingredient. The present invention also provides a use of the ginsenoside composition in the manufacture of a medicament for improving the activity of hair follicular tissue. By directly applying or spraying the medicament to the scalp, hair follicles in the telogen phase are encouraged to enter the regeneration cycle, thereby promoting hair growth. In addition, the present invention also provides a use of the ginsenoside composition in the manufacture of a medicament for improving the activity of in vitro-cultured hair follicular tissue. By preparing the ginsenoside composition as a tissue culture medium and applying it to in vitro-cultured hair follicular tissue, the activity of the cultured hair follicular tissue can be improved, significantly improving the survival rate of transplanted hair follicles.

[0005] CN112546056A discloses a composition for treating chemotherapy-induced peripheral neuropathy and its application, which relates to the field of medicine. The composition for treating chemotherapy-induced peripheral neuropathy provided by the present invention comprises ginsenoside Rg1 and fucoidan. The composition of the present invention significantly suppresses the expression of pro-inflammatory factors such as TNF-α, IL-1β, and IL-6, thereby exerting anti-inflammatory and repair-promoting effects on peripheral nerves and enhancing their resistance. Furthermore, the composition of the present invention significantly improves microcirculatory disorders and enhances SR-A expression, thereby enhancing the phagocytosis and clearance of HMGB1 by macrophages and suppressing the overexpression of TF. This reduces chemotherapy-induced microcirculatory disorders and neuralgia through multiple targets and mechanisms, fundamentally inhibiting the progression of CIPN, thereby achieving safe and effective treatment goals and providing a new therapeutic strategy for clinical practice.

[0006] The applicant has discovered through research that different combinations of ginsenosides (original ginseng saponin, diol-type ginsenosides, triol-type ginsenosides, secondary diol-type ginsenosides, and secondary triol-type ginsenosides) have different effects on the proliferation of epidermal keratinocytes and dermal papilla cells, as well as on the activity of 5α-reductase and alkaline phosphatase. This means that different combinations have different effects in terms of cytotoxicity, hair loss prevention, and hair growth and care.

[0007] The technical problem that this solution aims to solve is how to combine different ginsenosides to achieve optimal hair loss prevention and hair growth effects with the least amount of inactive ingredients. Summary of the Invention

[0008] In view of the shortcomings of the prior art, the object of the present invention is to provide a ginsenoside composition which can achieve optimal hair loss prevention and hair growth effects at a relatively low dose, ultimately resulting in a noticeable reduction in hair loss or thicker hair for the user.

[0009] At the same time, the present invention also provides uses of the composition and daily chemical products using the composition.

[0010] In order to achieve the object of this invention, the present invention adopts the following technical solution: A ginsenoside composition acting on hair, said ginsenoside composition being any one of ginsenoside compositions N1, N2, N4, N5 and N6, or a combination of multiple ginsenoside compositions; The N1 comprises the following components: Ginsenoside Rg1 12~28wt%, Ginsenoside Re 10-16 wt%, Ginsenoside Rb1 12~28wt%, Ginsenoside Rc 13~22wt%, Ginsenoside Rb2 13-22wt%, Ginsenoside Rd 7~13wt%, The N2 comprises the following components: Ginsenoside Rg1 30-50wt%, Ginsenoside Re 48~72wt%, The N4 comprises the following components: 20(S)-ginsenoside Rg2 8~16wt%, 20(R)-ginsenoside Rg2 7~15wt%, 20(S)-ginsenoside Rh1 17~25wt%, 20(R)-ginsenoside Rh1 16~23wt%, Ginsenoside Rk3 5-12 wt% Ginsenoside Rh4 16~26wt%, The N5 comprises the following ingredients: Ginsenoside Rk3 20-40wt%, Ginsenoside Rh4 60~80wt%, The N6 comprises the following components: 20(S)-ginsenoside Rg3 16-21wt%, 20(R)-ginsenoside Rg3 32~50wt%, Ginsenoside Rk1 4~8wt%, Ginsenoside Rg5 8-15wt%, 20(S)-ginsenoside Rh2 3~7wt%, 20(R)-ginsenoside Rh2 11-19wt%, Ginsenoside Rk2 0.1-1 wt% Ginsenoside Rh3 0.5-1.5wt%.

[0011] In the above-mentioned hair acting composition, N1 comprises the following ingredients: Ginsenoside Rg1 18-22wt%, Ginsenoside Re 12-15wt%, Ginsenoside Rb1 18-22wt%, Ginsenoside Rc 15-19 wt%, Ginsenoside Rb2 16-20wt%, Ginsenoside Rd 9~11wt%, The N2 comprises the following components: Ginsenoside Rg1 35-45wt%, Ginsenoside Re 55-65wt%, The N4 comprises the following components: 20(S)-ginsenoside Rg2 10-13 wt% 20(R)-ginsenoside Rg2 8-13 wt% 20(S)-ginsenoside Rh1 18~23wt%, 20(R)-ginsenoside Rh1 17~21wt%, Ginsenoside Rk3 6-10wt%, Ginsenoside Rh4 18~22wt%, The N5 comprises the following ingredients: Ginsenoside Rk3 25-35wt%, Ginsenoside Rh4 65~75wt%, The N6 comprises the following components: 20(S)-ginsenoside Rg3 18-20wt%, 20(R)-ginsenoside Rg3 37~43wt%, Ginsenoside Rk1 5-7wt%, Ginsenoside Rg5 9-13 wt% 20(S)-ginsenoside Rh2 4~6wt%, 20(R)-ginsenoside Rh2 13~17wt%, Ginsenoside Rk2 0.2~0.8wt%, Ginsenoside Rh3 0.7~1.2wt%.

[0012] The above-mentioned ginsenoside composition acting on hair includes at least one of N1, N2, and N4, and at least one of N4, N5, and N6.

[0013] In a preferred embodiment of the present invention, the ginsenoside composition contains 0.1 to 99.9 wt% of at least one of N1, N2, and N4, and 0.1 to 99.9 wt% of at least one of N4, N5, and N6: In a further preferred embodiment of the present invention, the composition contains 1 to 99 wt% of at least one of N1, N2, and N4, and 1 to 99 wt% of at least one of N4, N5, and N6.

[0014] In a further preferred embodiment of the present invention, the composition contains 10-90 wt% of at least one of N1, N2, and N4, and 10-90 wt% of at least one of N4, N5, and N6: In a further preferred embodiment of the present invention, the composition contains 20-80 wt% of at least one of N1, N2, and N4, and 20-80 wt% of at least one of N4, N5, and N6: In a further preferred embodiment of the present invention, the composition contains 30-70 wt% of at least one of N1, N2, and N4, and 30-70 wt% of at least one of N4, N5, and N6: In a further preferred embodiment of the present invention, the composition contains 40-60 wt% of at least one of N1, N2, and N4, and 40-60 wt% of at least one of N4, N5, and N6: In a further preferred embodiment of the invention, the composition comprises 50 wt% of at least one of N1, N2, N4 and 50 wt% of at least one of N4, N5, N6. At the same time, the present invention further discloses a daily chemical product acting on hair, which contains any of the above-described ginsenoside compositions. The concentration of the ginsenoside composition in the daily chemical product is 0.05 to 10000 μg / ml, preferably, the concentration of the composition in the daily chemical product is 1 to 5000 μg / ml.

[0015] Preferably, the concentration of the ginsenoside composition in the daily chemical product is 10-1000 μg / ml.

[0016] Although the concentration used in the cell experiments of the present invention is relatively low, when applied to daily formulations, it needs to be 10 or 100 times the cell concentration.

[0017] In the above-mentioned daily chemical products acting on hair, the daily chemical products include shampoo, conditioner, freeze-dried powder, essence, shampoo spray, hair care cream, hair care milk, etc.

[0018] At the same time, the present invention further discloses the use of ginsenoside composition as an active ingredient for preventing hair loss in daily chemical products. The ginsenoside composition is N1, N2 or N4, The N1 comprises the following components: Ginsenoside Rg1 12~28wt%, Ginsenoside Re 10-16 wt%, Ginsenoside Rb1 12~28wt%, Ginsenoside Rc 13~22wt%, Ginsenoside Rb2 13-22wt%, Ginsenoside Rd 7~13wt%, The N2 comprises the following components: Ginsenoside Rg1 30-50wt%, Ginsenoside Re 48~72wt%, The N4 comprises the following components: 20(S)-ginsenoside Rg2 8~16wt%, 20(R)-ginsenoside Rg2 7~15wt%, 20(S)-ginsenoside Rh1 17~25wt%, 20(R)-ginsenoside Rh1 16~23wt%, Ginsenoside Rk3 5-12 wt% Ginsenoside Rh4 16-26wt%.

[0019] Preferably, N1 comprises the following components: Ginsenoside Rg1 18-22wt%, Ginsenoside Re 12-15wt%, Ginsenoside Rb1 18-22wt%, Ginsenoside Rc 15-19 wt%, Ginsenoside Rb2 16-20wt%, Ginsenoside Rd 9~11wt%, The N2 comprises the following components: Ginsenoside Rg1 35-45wt%, Ginsenoside Re 55-65wt%, The N4 comprises the following components: 20(S)-ginsenoside Rg2 10-13 wt% 20(R)-ginsenoside Rg2 8-13 wt% 20(S)-ginsenoside Rh1 18~23wt%, 20(R)-ginsenoside Rh1 17~21wt%, Ginsenoside Rk3 6-10wt%, Ginsenoside Rh4 18-22wt%.

[0020] At the same time, the present invention further discloses the use of the ginsenoside composition as an active ingredient for hair growth and hair regrowth in daily chemical products. The ginsenoside composition is N4, N5 or N6, The N4 comprises the following components: 20(S)-ginsenoside Rg2 8~16wt%, 20(R)-ginsenoside Rg2 7~15wt%, 20(S)-ginsenoside Rh1 17~25wt%, 20(R)-ginsenoside Rh1 16~23wt%, Ginsenoside Rk3 5-12 wt% Ginsenoside Rh4 16~26wt%, The N5 comprises the following ingredients: Ginsenoside Rk3 20-40wt%, Ginsenoside Rh4 60~80wt%, The N6 comprises the following components: 20(S)-ginsenoside Rg3 16-21wt%, 20(R)-ginsenoside Rg3 32~50wt%, Ginsenoside Rk1 4~8wt%, Ginsenoside Rg5 8-15wt%, 20(S)-ginsenoside Rh2 3~7wt%, 20(R)-ginsenoside Rh2 11-19wt%, Ginsenoside Rk2 0.1-1 wt% Ginsenoside Rh3 0.5~1.5wt%, Preferably, the N4 comprises the following components: 20(S)-ginsenoside Rg2 10-13 wt% 20(R)-ginsenoside Rg2 8-13 wt% 20(S)-ginsenoside Rh1 18~23wt%, 20(R)-ginsenoside Rh1 17~21wt%, Ginsenoside Rk3 6-10wt%, Ginsenoside Rh4 18~22wt%, The N5 comprises the following ingredients: Ginsenoside Rk3 25-35wt%, Ginsenoside Rh4 65~75wt%, The N6 comprises the following components: 20(S)-ginsenoside Rg3 18-20wt%, 20(R)-ginsenoside Rg3 37~43wt%, Ginsenoside Rk1 5-7wt%, Ginsenoside Rg5 9-13 wt% 20(S)-ginsenoside Rh2 4~6wt%, 20(R)-ginsenoside Rh2 13~17wt%, Ginsenoside Rk2 0.2~0.8wt%, Ginsenoside Rh3 0.7~1.2wt%.

[0021] In the above, the total of each component in N1, N2, N4, N5, N6, and N7 is 100%.

[0022] Compared with the prior art, the present invention has the following beneficial effects: The present inventors conducted experiments using various combinations of the proliferation of epidermal keratinocytes and dermal papilla cells, 5α-reductase activity, and alkaline phosphatase activity, and clarified the following. The cytotoxicity of ginsenoside composition N5, ginsenoside composition N6, and ginsenoside composition N7 is stronger than that of other ginseng extracts.

[0023] Ginsenoside composition N4 has the effect of promoting the proliferation of epidermal keratinocytes, and in a test using a bilayer culture system of dermal papilla cells and epidermal keratinocytes, it was also found to have the effect of promoting the proliferation of epidermal keratinocytes.

[0024] Ginsenoside Composition N1, Ginsenoside Composition N2, and Ginsenoside Composition N4 have the effect of inhibiting the activity of 5α-reductase.

[0025] Ginsenoside Composition N4, Ginsenoside Composition N5, and Ginsenoside Composition N6 have the effect of promoting the activity of alkaline phosphatase in cultured hair papilla cells.

[0026] This means that ginsenoside composition N1, ginsenoside composition N2, and ginsenoside composition N4 have the effect of preventing male pattern baldness, while ginsenoside composition N4, ginsenoside composition N5, and ginsenoside composition N6 have the effect of promoting hair growth. In summary, ginsenoside composition N4 showed the best performance in most tests and is considered to be the best choice for adding to daily chemical products as the best option for preventing hair loss and promoting hair growth. [Brief explanation of the drawings]

[0027] [Figure 1] 1 is a graph showing the effect of ginsenoside composition on the proliferation of epidermal keratinocytes. Mean value + standard deviation n=3, + indicates P<0.1, * indicates P<0.05, ** indicates P<0.01 vs. control, and the same applies below. [Figure 2] 1 is a graph showing the effect of a ginsenoside composition on the proliferation of dermal papilla cells. [Figure 3] 1 is a graph showing the effect of a carrot extract on the proliferation of epidermal keratinocytes in a bilayer culture system of dermal papilla cells and keratinocytes when the concentration of the ginsenoside composition is 2 μg / ml. [Figure 4] 1 is a graph showing the effect of a carrot extract on the proliferation of epidermal keratinocytes in a bilayer culture system of dermal papilla cells and keratinocytes when the concentration of the ginsenoside composition is 20 μg / ml. [Figure 5] 1 is a graph showing the effect of carrot extract on the proliferation of epidermal keratinocytes in a bilayer culture system of dermal papilla cells and keratinocytes at different concentrations of minoxidil sulfate. [Figure 6] 1 is a graph showing the effect of ginsenoside compositions on 5α-reductase activity. [Figure 7] 1 is a graph showing the effect of N4 sets of ginsenoside compositions on 5α-reductase activity. [Figure 8] FIG. 1 shows a staining image of alkaline phosphatase activity in cultured dermal papilla cells when the concentration of a ginsenoside composition is 2 μg / ml. [Figure 9] FIG. 1 is a staining diagram of alkaline phosphatase activity in cultured dermal papilla cells when the concentration of a ginsenoside composition is 20 μg / ml. [Figure 10] 1 is a graph showing the effect of ginsenoside compositions on alkaline phosphatase activity of cultured dermal papilla cells. [Figure 11] 1 is a graph showing the effect of different concentrations of valproic acid on alkaline phosphatase activity of cultured dermal papilla cells, with valproic acid used as a positive control. [Figure 12] These are the test results for test report number G522-2210027. [Figure 13] These are the test results for test report number G522-2209047. [Figure 14] This is a microscopic photograph of the scalp of some subjects in a human study. [Figure 15] This is a photograph of the hair of some subjects in a human study. DETAILED DESCRIPTION OF THE INVENTION

[0028] The technical solutions of the present invention will be further described below with reference to specific embodiments. Those skilled in the art should understand that the above examples are only intended to help understand the present invention and should not be considered as specifically limiting the present invention.

[0029] Unless specific experimental procedures or conditions are specified in the examples, they can be performed according to the operations and conditions of conventional experimental procedures described in the literature in the field. Unless the manufacturers of the reagents and equipment used are specified, they are all conventional reagent products available commercially. Example 1

[0030] A ginsenoside composition containing the ingredients shown in Table 1 below: JPEG2026502711000002.jpg105170 (Example 2)

[0031] A ginsenoside composition containing the ingredients shown in Table 2 below: JPEG2026502711000003.jpg63170 (Example 3)

[0032] The ginsenoside composition (N3) contained the following ingredients: Ginsenoside Rb1 30wt%, Ginsenoside Rc 26wt%, Ginsenoside Rb2 27wt%, Ginsenoside Rd 17wt%. Example 4

[0033] A ginsenoside composition containing the ingredients shown in Table 3 below: JPEG2026502711000004.jpg86170 (Example 5)

[0034] A ginsenoside composition containing the ingredients shown in Table 4 below: JPEG2026502711000005.jpg61170 (Example 6)

[0035] A ginsenoside composition containing the ingredients shown in Table 5 below: JPEG2026502711000006.jpg118170 (Example 7)

[0036] A ginsenoside composition comprising the following ingredients: Ginsenoside Rk1 13wt%, 20(S)-ginsenoside Rh2 23wt%, 20(R)-ginsenoside Rh2 13wt%, Ginsenoside Rk2 20wt%, Ginsenoside Rh3 31wt%.

[0037] Preparation methods for Examples 1 to 7 Accurately weigh and blend each ingredient. Performance Test

[0038] Ginsenoside compositions N1, N2, N3, N4, N5, N6, and N7 were dissolved in dimethyl sulfoxide (DMSO) to prepare samples with concentrations of 20 mg / ml, 2 mg / ml, 0.2 mg / ml, and 0.02 mg / ml, respectively. As a positive control, minoxidil sulfate (Sigma-Aldrich) was dissolved in DMSO to prepare solutions with concentrations of 2 mg / ml, 0.2 mg / ml, 0.02 mg / ml, and 0.002 mg / ml.

[0039] At the same time, finasteride (Sigma-Aldrich) was also prepared in DMSO to a concentration of 1.5 μg / ml (4 μM). 1. Effect of carrot extract on the proliferation of epidermal keratinocytes and dermal papilla cells A human epidermal keratinocyte solution at a concentration of 200,000 cells / cell was seeded onto a 96-well cell culture plate (n=3) at 100 μl / well and cultured for 1 day. Next, 1 μl / well of carrot extracts 1–7, a positive control (minoxidil sulfate), and a blank control (DMSO) were added, and the plates were cultured for 3 days. Then, 10 μl / well of Cell Counting Kit-8 (Dojindo Laboratories) was added, and the plates were cultured for an additional 2 hours. The cell count was measured using a microplate reader (450 nm, Mix 0).

[0040] For the effect of ginsenoside composition on the proliferation of epidermal keratinocytes, see FIG. In FIG. 1, ginsenoside composition N4 significantly increased the cell number at concentrations of 0.2 μg / ml and 2 μg / ml (p values: P<0.05, P<0.1, respectively).

[0041] The effect was higher than that of the positive control minoxidil sulfate (0.02μg / ml, 0.2μg / ml, 2μg / ml, 20μg / ml). At the same time, ginsenoside compositions N4, N5, N6, and N7 significantly reduced cell counts at a concentration of 200μg / ml. Ginsenoside composition N1 also reduced cell counts at a concentration of 200μg / ml. A human dermal papilla cell solution with a concentration of 100,000 cells / cell was seeded into a 96-well cell culture plate (n=3) at 100 μl / well and cultured for one day. The proliferation and counting of dermal papilla cells were performed in the same manner. The concentrations of the test samples ranged from 0.2 μg / ml to 200 μg / ml, and the concentrations of minoxidil sulfate ranged from 0.02 μg / ml to 20 μg / ml.

[0042] Referring to FIG. 2, FIG. 2 shows the effect of ginsenoside compositions on the proliferation of dermal papilla cells. Ginsenoside composition N4 has the effect of increasing the cell number at a concentration of 2 μg / ml (P<0.05).

[0043] This effect was greater than that of the positive control minoxidil sulfate (0.02 μg / ml, 0.2 μg / ml, 2 μg / ml, 20 μg / ml). Ginsenoside composition N3 also had the effect of increasing cell number at a concentration of 2 μg / ml (P<0.05). Ginsenoside composition N2 tended to increase cell number at a concentration of 0.2 μg / ml (P<0.1). Ginsenoside compositions N4 and N6 significantly reduced cell number at a concentration of 200 μg / ml. Ginsenoside compositions N1, 3, 5, and 7 also reduced cell number at 200 μg / ml.

[0044] This method uses a highly sensitive water-soluble yellow formazan product to generate a water-soluble tetrazolium salt, which is reduced by intracellular dehydrogenases. The absorbance of the resulting water-soluble yellow formazan product at 450 nm is measured to calculate the number of viable cells. There is a linear relationship between the number of cells and the amount of formazan.

[0045] 2. Effect of carrot extract on proliferation of epidermal keratinocytes in a bilayer culture system of dermal papilla cells and epidermal keratinocytes 500 μl of human epidermal keratinocytes (30,000 cells / ml) were added to each well of a 24-well MULTIWELL plate, followed by a 1.0 μm cell culture insert and incubation for 1 day. 300 μl of dermal papilla cells (20,000 cells / ml) were added to the 1.0 μm cell culture insert. Carrot extracts 1–7 were then added, followed by 1 μl / well of minoxidil sulfate as a positive control and a blank control (DMSO), and incubation continued for 2 days. The 1.0 μm cell culture insert was removed, and 50 μl / well of Cell Counting Kit-8 (Dojindo Laboratories) was added, followed by incubation for 2 hours. Cell counts were measured using a microplate reader (450 nm, Mix 0).

[0046] Referring to FIG. 3, FIG. 3 shows the effect of a carrot extract on the proliferation of epidermal keratinocytes in a bilayer culture system of dermal papilla cells and keratinocytes when the concentration of the ginsenoside composition is 2 μg / ml. Referring to FIG. 4, FIG. 3 shows the effect of ginseng extract on the proliferation of epidermal keratinocytes in a bilayer culture system of dermal papilla cells and keratinocytes when the concentration of the ginsenoside composition is 20 μg / ml. Referring to FIG. 5, FIG. 5 is a graph showing the effect of carrot extract on the proliferation of epidermal keratinocytes in a bilayer culture system of dermal papilla cells and keratinocytes at different concentrations of minoxidil sulfate. In a test using a bilayer culture system of dermal papilla cells and keratinocytes, ginsenoside compositions N4 and 7 were found to have the effect of promoting keratinocyte proliferation at a concentration of 2 μg / ml. Ginsenoside compositions N5, 6, and 7 had inhibitory or cytotoxic effects at a concentration of 20 μg / ml. The positive control minoxidil sulfate (0.2 μg / ml, 2 μg / ml, 20 μg / ml, 200 μg / ml) was not found to have the effect of promoting keratinocyte proliferation.

[0047] 3. Effect of carrot extract on 5α-reductase activity 5α-reductase is an enzyme that promotes the conversion of testosterone to 5α-dihydrotestosterone. This study measured the effect of carrot extract on 5α-reductase activity using the NAD cycling method. Solution A was prepared by adding 520 μl of a 0.3 M sucrose / 1 mM dithiothreitol / 40 mM potassium phosphate buffer solution (pH 5.0), 80 μl of 100 μM testosterone, and 40 μl of 16 μM NADPH to a 1.5 ml test tube. 83 μl of solution A, 1 μl of carrot extract, and 16 μl of rat liver microsomes were added and incubated for 20 minutes. After heating at 80°C for 5 minutes, 600 μl of Tris-HCl buffer solution (pH 9.8) and 40 μl of 20 mM thio-NAD were added and incubated for 10 minutes. After centrifugation, 100 μl of the solution was added to a microplate, followed by the addition of 10 μl of 400 U / ml 3α-HSD. The absorbance was measured at 400 nm within 30 minutes. Finasteride (40 nM) served as a positive control.

[0048] FIG. 6 is a graph showing the effect of ginsenoside compositions on 5α-reductase activity. Ginsenoside compositions N1, 2, and 4 (200 μg / ml) have an inhibitory effect on the activity of 5α-reductase, the strength of which is equivalent to that of 15 ng / ml (40 nM) finasteride.

[0049] 7, ginsenoside compositions N4, N4-1, N4-2, N4-3, N4-4, and N4-5 (200 μg / ml) have an inhibitory effect on the activity of 5α-reductase. The strength of the inhibitory effect is almost the same.

[0050] 4. Effect of carrot extract on alkaline phosphatase activity A human dermal papilla cell solution was prepared at a concentration of 100,000 cells / ml and added to a 96-well cell culture plate at 100 μl / well (n=3). The cells were cultured for one day. The test sample concentrations ranged from 0.2 μg / ml to 200 μg / ml, and the minoxidil sulfate concentrations ranged from 0.02 μg / ml to 20 μg / ml. After three days of culture, the cells were fixed with 4% PFA, washed with PBS, and then alkaline phosphatase activity was measured at 405 nm using BCIP-NBT solution.

[0051] 8 is a staining image of alkaline phosphatase activity of cultured dermal papilla cells when the concentration of the ginsenoside composition was 2 μg / ml. Valproic acid was used as a positive control. 9 is a staining image of alkaline phosphatase activity of cultured dermal papilla cells when the concentration of the ginsenoside composition was 20 μg / ml. Valproic acid was used as a positive control. FIG. 10 is a graph showing the effect of ginsenoside compositions on alkaline phosphatase activity of cultured dermal papilla cells. FIG. 11 is a graph showing the effect of different concentrations of valproic acid on alkaline phosphatase activity of cultured dermal papilla cells, using valproic acid as a positive control.

[0052] Analyzing the results: 1. The cytotoxicity of ginsenoside composition N5, ginsenoside composition N6, and ginsenoside composition N7 was stronger than that of other ginseng extracts (see Figure 4).

[0053] 2. Referring to Figures 1 to 4, ginsenoside composition N4 has the effect of promoting the proliferation of epidermal keratinocytes, and also showed the effect of promoting the proliferation of epidermal keratinocytes when tested using a bilayer culture system of dermal papilla cells and epidermal keratinocytes.

[0054] 3. Referring to Figure 6, ginsenoside composition N1, ginsenoside composition N2, and ginsenoside composition N4 have the effect of inhibiting the activity of 5α-reductase, suggesting that N1, N2, and N4 can significantly prevent hair loss in men.

[0055] 4. Referring to Figure 7, ginsenoside compositions N4, N4-1, N4-2, N4-3, N4-4, and N4-5 have almost the same inhibitory effect on 5α-reductase activity, suggesting that N4, N4-1, N4-2, N4-3, N4-4, and N4-5 can all effectively and significantly prevent male hair loss.

[0056] 5. Referring to Figures 8 to 10, ginsenoside composition N4, ginsenoside composition N5, and ginsenoside composition N6 have a promoting effect on the alkaline phosphatase activity of cultured hair papilla cells, suggesting that N4, N5, and N6 have the effect of promoting hair growth and increasing hair volume.

[0057] In summary, N1, N2, and N4 have the effect of preventing male pattern baldness, N4, N5, and N6 have the effect of promoting hair growth and increasing hair volume, and N5, N6, and N7 have cytotoxicity. N4 is the ingredient with the greatest potential for commercial application from the perspective of promoting the proliferation of epidermal keratinocytes, preventing hair loss, promoting hair growth, and increasing hair volume.

[0058] For a more effective approach, it is recommended to combine N1, N2, or N4 with N4, N5, or N6. For a more effective method, it is recommended to combine N1 or N2 with N4.

[0059] The N4 formulation of the present invention, under the trade name "Sanshi@ye SPC," was sent to Shaanxi Boxi General Testing Technology Co., Ltd. for an in vitro hair loss prevention test. N4 was the only active ingredient; the other ingredients were auxiliary solvents and preservatives without hair loss prevention or hair growth functions. The test was divided into four parts. In Part 1, cytotoxicity was detected using dermal papilla cells to determine the safe administration concentration of the sample on dermal papilla cells. In Part 2, dihydrotestosterone (DHT) was used to stimulate dermal papilla cells, and the hair loss prevention and hair growth effects of the test sample were evaluated by detecting changes in VEGF content and DKK1 gene expression after the sample was applied to dermal papilla cells. In Part 3, the hair growth effects of the test sample were evaluated by detecting cell proliferation after the sample was applied to dermal papilla cells. In Part 4, the hair loss prevention effects of the test sample were evaluated by inhibiting 5α-reductase activity in vitro.

[0060] Test report number: G522-2210027, test period: August 25th to September 25th, 2022, please refer to Figure 12 for test results. Test report number: G522-2209047, test period: August 24th to September 4th, 2022, please refer to Figure 13 for test results.

[0061] The ingredients of the raw material "Sanshi R Night SPC" mentioned in the above two test reports are as shown in Table 6 below. JPEG2026502711000007.jpg57170The above test results also confirmed that the addition of N4 has better hair loss prevention and hair growth effects.

[0062] At the same time, the above-mentioned "Sanshi R Night SPC" was added to a regular essence (see Recipe Table 7) to prepare an anti-hair loss essence. Among these, Sanshi R Night SPC was the only active ingredient; the other ingredients were auxiliary solvents and preservatives without any hair loss prevention or hair growth function. The essence was sent to Hua Measuring Testing and Certification Group Co., Ltd. for human testing. The sample number was HBO00942001, and the experiment was divided into three sections: hair loss count test, overall hair density test, and local skin hair test. JPEG2026502711000008.jpg127170 Process: Water was added to the water pot, the carbomer was slowly added, stirring was started, and the temperature was raised to 85°C. The water was suctioned from the water pot into the main pot, and a premix of butanediol and sodium hyaluronate was added, maintained at 85°C, and stirred uniformly. The mixture was cooled to 60°C, and the aqueous solution of phase B was added and stirred uniformly. After cooling to 40°C, the premixed liquid of phase C was added and stirred uniformly, and then phase D was added and stirred uniformly. The materials were collected and tested, and those that met the pH standard of 5.5-6.5 were selected.

[0063] Physical and chemical parameters: Appearance: transparent essence-like liquid pH: 5.5~6.5 See Table 8 for hair loss count test results. JPEG2026502711000009.jpg130170As can be seen from the above test, after 10 subjects used the sample for 28 days, the hair loss count decreased by 9.52%, after using the sample for 56 days, the hair loss count decreased by 38.10%, and after using the sample for 84 days, the hair loss count decreased by 66.67%. During the product use period, the subjects' hair loss situation improved significantly with increasing use time.

[0064] See Table 9 for overall hair density testing. JPEG2026502711000010.jpg130170As can be seen from the above tests, with increasing usage time, after 10 subjects used the sample for 28 days, the overall hair density increased by 5.26%, after using the sample for 56 days, the overall hair density increased by 13.16%, and after using the sample for 84 days, the overall hair density increased by 18.42%.

[0065] Figure 14 shows micrographs of the scalp of some subjects. Figure 15 shows photographs of the hair of some of the subjects.

[0066] As can be seen from human tests, when the composition N4 of the present invention is used as an essence, it can significantly inhibit hair loss and improve hair density.

[0067] Below, we provide examples of formulations in which the composition (N4) of the present invention is applied to other representative scalp formulations. The ginsenosides in the following tables are composition (N4) as follows: The JPEG2026502711000011.jpg169170 process is as follows: 1. Phases A and B: Heat the water to 85°C, add Phases A and B, and stir to dissolve.

[0068] 2. Phases C and D: Cool to 40°C, add phases C and D, and stir evenly.

[0069] 3. Phase F was added, the pH was adjusted, and the mixture was stirred uniformly.

[0070] 4. The stirring was stopped and the material was removed.

[0071] Physical and chemical parameters: Appearance: Clear viscous liquid.

[0072] Viscosity: (25℃, 3#6rpm): 5000~12000mPa·s, pH (20℃, 10% aqueous solution): 6.2~6.4

[0073] The JPEG2026502711000012.jpg182170 process is as follows: 1. Phase A: Heat to 85°C and stir to dissolve.

[0074] 2. Phases B, C, D, and E: Cooled to 40°C and then added. SF-1 was pre-dispersed and dissolved in water, then added to the materials and stirred evenly.

[0075] 3. The stirring was stopped and the material was removed.

[0076] Physical and chemical parameters: Appearance: green particles and transparent liquid, Viscosity: (25℃, 3#6rpm): 9000~12000mPa·s, pH (20℃, 10% aqueous solution): 6.5~6.8 JPEG2026502711000013.jpg59170The above test reveals the following: When different ginsenosides were used in combination, the combination of triol-based rare ginsenosides with secondary saponins (N4 and N5), the combination of diol-based rare ginsenosides with secondary saponins (N6, N7), the mixed original ginsenoside (N1), diol-based ginsenoside (N2), and triol-based ginsenoside (N3) showed different trends. Among them, N4 showed the best effect in multiple tests and is a combination worthy of continued research. The salient contributions of the present invention are as follows: 1. Different ginsenosides have different properties, so when using them, they should be flexibly selected and formulated according to the characteristics of the product formulation and based on the above experimental results.

[0077] 2. The present invention optimizes raw materials as much as possible and provides some combinations with the least amount of raw materials to be applied to the formulation of daily chemical products with different functions.

Claims

1. A ginsenoside composition acting on hair, the composition being any one of ginsenoside compositions N1, N2, N4, N5 and N6, or a combination of multiple ginsenoside compositions; The N1 comprises the following components: ginsenoside Rg1 12-28 wt%, ginsenoside Re 10-16 wt%, ginsenoside Rb1 12-28 wt%, ginsenoside Rc 13-22 wt%, ginsenoside Rb2 13-22 wt%, Ginsenoside Rd 7-13 wt%, The N2 comprises the following components: ginsenoside Rg1 30-50 wt%; ginsenoside Re 48-72 wt%, The N4 comprises the following components: 20(S)-ginsenoside Rg2 8-16 wt% 20(R)-ginsenoside Rg2 7 to 15 wt %, 20(S)-ginsenoside Rh1 17-25 wt %, 20(R)-ginsenoside Rh1 16-23 wt%, ginsenoside Rk3 5-12 wt%; ginsenoside Rh4 16-26 wt%, The N5 comprises the following components: ginsenoside Rk3 20-40 wt%; ginsenoside Rh4 60-80 wt%, The N6 comprises the following components: 20(S)-ginsenoside Rg3 16-21 wt%; 20(R)-ginsenoside Rg3 32-50 wt%; ginsenoside Rk1 4-8 wt%, ginsenoside Rg5 8-15 wt%, 20(S)-ginsenoside Rh2 3-7 wt%; 20(R)-ginsenoside Rh2 11 to 19 wt %, ginsenoside Rk2 0.1 to 1 wt%; ginsenoside Rh3 0.5 to 1.5 wt%; A ginsenoside composition acting on hair, characterized in that the total amount of each component in N1, N2, N4, N5, N6 and N7 is 100%.

2. The N1 comprises the following components: Ginsenoside Rg1 18-22 wt%, ginsenoside Re 12-15 wt%, ginsenoside Rb1 18-22 wt%, ginsenoside Rc 15-19 wt%, ginsenoside Rb2 16-20 wt%; Ginsenoside Rd 9-11 wt%, The N2 comprises the following components: ginsenoside Rg1 35-45 wt%, ginsenoside Re 55-65 wt%, The N4 comprises the following components: 20(S)-ginsenoside Rg2 10-13 wt% 20(R)-ginsenoside Rg2 8 to 13 wt%; 20(S)-ginsenoside Rh1 18-23 wt%, 20(R)-ginsenoside Rh1 17-21 wt%, ginsenoside Rk3 6-10 wt%; ginsenoside Rh4 18-22 wt%, The N5 comprises the following components: ginsenoside Rk3 25-35 wt%, ginsenoside Rh4 65-75 wt%, The N6 comprises the following components: 20(S)-ginsenoside Rg3 18-20 wt%; 20(R)-ginsenoside Rg3 37-43 wt%, ginsenoside Rk1 5-7 wt%, ginsenoside Rg5 9-13 wt%; 20(S)-ginsenoside Rh2 4-6 wt% 20(R)-ginsenoside Rh2 13 to 17 wt %, ginsenoside Rk2 0.2 to 0.8 wt%; ginsenoside Rh3 0.7 to 1.2 wt%; 2. The ginsenoside composition acting on hair according to claim 1, wherein the total of each component in N1, N2, N4, N5, N6 and N7 is 100%.

3. 2. The ginsenoside composition acting on hair according to claim 1, characterized in that it contains at least one of N1, N2, and N4, and at least one of N4, N5, and N6.

4. A daily chemical product acting on hair, comprising the ginsenoside composition according to any one of claims 1 to 3, A daily chemical product acting on hair, characterized in that the concentration of the composition in the daily chemical product is 0.05 to 10,000 μg / ml, preferably the concentration of the composition in the daily chemical product is 1 to 5,000 μg / ml.

5. 5. The hair-care product according to claim 4, wherein the concentration of the ginsenoside composition in the hair care product is 10 to 1000 μg / ml.

6. The hair-care daily chemical product according to claim 4, characterized in that the hair-care daily chemical product is a shampoo, a conditioner, a freeze-dried powder, an essence, a shampoo spray, a hair care cream, or a hair care milk.

7. Use of a ginsenoside composition as an active ingredient for preventing hair loss in daily chemical products, wherein the ginsenoside composition is N1, N2 or N4; The N1 comprises the following components: ginsenoside Rg1 12-28 wt%, ginsenoside Re 10-16 wt%, ginsenoside Rb1 12-28 wt%, ginsenoside Rc 13-22 wt%, ginsenoside Rb2 13-22 wt%, Ginsenoside Rd 7-13 wt%, The N2 comprises the following components: ginsenoside Rg1 30-50 wt%; ginsenoside Re 48-72 wt%, The N4 comprises the following components: 20(S)-ginsenoside Rg2 8-16 wt% 20(R)-ginsenoside Rg2 7 to 15 wt %, 20(S)-ginsenoside Rh1 17-25 wt %, 20(R)-ginsenoside Rh1 16-23 wt%, ginsenoside Rk3 5-12 wt%; ginsenoside Rh4 16-26 wt%, A use characterized in that the total of each component in N1, N2, and N4 is 100%.

8. The N1 comprises the following components: Ginsenoside Rg1 18-22 wt%, ginsenoside Re 12-15 wt%, ginsenoside Rb1 18-22 wt%, ginsenoside Rc 15-19 wt%, ginsenoside Rb2 16-20 wt%; Ginsenoside Rd 9-11 wt%, The N2 comprises the following components: ginsenoside Rg1 35-45 wt%, ginsenoside Re 55-65 wt%, The N4 comprises the following components: 20(S)-ginsenoside Rg2 10-13 wt% 20(R)-ginsenoside Rg2 8 to 13 wt%; 20(S)-ginsenoside Rh1 18-23 wt%, 20(R)-ginsenoside Rh1 17-21 wt%, ginsenoside Rk3 6-10 wt%; ginsenoside Rh4 18-22 wt%, The use according to claim 7, characterized in that the sum of each component in N1, N2 and N4 is 100%.

9. Use of a ginsenoside composition as an active ingredient for hair growth and hair regrowth in daily chemical products, wherein the ginsenoside composition is N4, N5 or N6; The N4 comprises the following components: 20(S)-ginsenoside Rg2 8-16 wt% 20(R)-ginsenoside Rg2 7 to 15 wt %, 20(S)-ginsenoside Rh1 17-25 wt %, 20(R)-ginsenoside Rh1 16-23 wt%, ginsenoside Rk3 5-12 wt%; ginsenoside Rh4 16-26 wt%, The N5 comprises the following components: ginsenoside Rk3 20-40 wt%; ginsenoside Rh4 60-80 wt%, The N6 comprises the following components: 20(S)-ginsenoside Rg3 16-21 wt%; 20(R)-ginsenoside Rg3 32-50 wt%; ginsenoside Rk1 4-8 wt%, ginsenoside Rg5 8-15 wt%, 20(S)-ginsenoside Rh2 3-7 wt%; 20(R)-ginsenoside Rh2 11 to 19 wt %, ginsenoside Rk2 0.1 to 1 wt%; ginsenoside Rh3 0.5 to 1.5 wt%; The total of each component in N4, N5, and N6 is 100%.

10. The N4 comprises the following components: 20(S)-ginsenoside Rg2 10-13 wt% 20(R)-ginsenoside Rg2 8 to 13 wt%; 20(S)-ginsenoside Rh1 18-23 wt%, 20(R)-ginsenoside Rh1 17-21 wt%, ginsenoside Rk3 6-10 wt%; ginsenoside Rh4 18-22 wt%, The N5 comprises the following components: ginsenoside Rk3 25-35 wt%, ginsenoside Rh4 65-75 wt%, The N6 comprises the following components: 20(S)-ginsenoside Rg3 18-20 wt%; 20(R)-ginsenoside Rg3 37-43 wt%, ginsenoside Rk1 5-7 wt%, ginsenoside Rg5 9-13 wt%; 20(S)-ginsenoside Rh2 4-6 wt% 20(R)-ginsenoside Rh2 13 to 17 wt %, ginsenoside Rk2 0.2 to 0.8 wt%; ginsenoside Rh3 0.7 to 1.2 wt%; The use according to claim 9, characterized in that the total of each component in N4, N5 and N6 is 100%.

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