A composition for preventing hair loss containing an extract of norway spruce leaves and use thereof

Abstract

The application discloses a hair loss prevention composition containing Norway spruce leaf extract and application, and relates to the technical field of cosmetics.The composition comprises component A and component B.The component A comprises the following components in parts by weight: 1-5 parts of verbascum thapsus extract, 0.1-1 parts of saw palmetto extract and 1-5 parts of Norway spruce leaf extract.The component B comprises the following components in parts by weight: 2-4 parts of eucommia ulmoides bark extract.The verbascum thapsus extract, the saw palmetto extract, the Norway spruce leaf extract and the eucommia ulmoides bark extract are combined to have a synergistic effect.The composition formed by the specific combination of the components can prevent androgenic alopecia by anti-inflammatory, antibacterial, inhibition of TRPV3 channel and inhibition of the activity of 5alpha-reductase, can inhibit the secretion of lipids by sebaceous cells, and can achieve the purpose of oil control, and the components are all plant components, so the composition is more suitable for men.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and in particular to an anti-hair loss composition containing Norway spruce leaf extract and its application. Background Technology

[0002] Common types of hair loss include androgenetic alopecia (seborrheic alopecia), immune-related alopecia (neurotic alopecia, alopecia areata), and alopecia areata caused by external stimuli. Androgenetic alopecia is an androgen-dependent, autosomal dominant, polygenic hereditary hair loss that often occurs alongside sebum secretion, hence the name seborrheic alopecia. Its mechanism involves testosterone being converted to the more potent dihydrotestosterone (DHT) by 5α-reductase. DHT then enters the cell nucleus, activating specific genes to produce specific proteins that regulate hair follicle cell proliferation and differentiation, shortening the anagen phase and causing premature regression. Currently, most cosmetics achieve hair loss prevention through anti-hormonal effects. For example, Polygonum multiflorum extract can reduce androgen production, and Ganoderma lucidum and Platycladus orientalis leaf extracts can inhibit the action of 5α-reductase. However, long-term use of hormone-inhibiting ingredients may disrupt the body's hormonal balance. Furthermore, since the scalp is connected to facial skin, scalp care may affect facial hair growth. For men, long-term use of hormone-inhibiting ingredients can also negatively impact secondary sexual characteristics. Therefore, suppressing androgens or 5α-reductase to reduce hair loss is not suitable for men, and alternative methods need to be explored.

[0003] Recent studies have shown that the TRPV3 channel is closely related to hair growth. TRPV3 is highly expressed in cells surrounding the hair follicle; activation of TRPV3 can inhibit the proliferation of outer root sheath cells, thereby suppressing hair growth and inducing keratinocyte apoptosis. The TRPV3 channel can be activated by certain activators (such as carvacrol and 2-APB) and at higher temperatures (≥33℃). For men, frequent exercise or outdoor physical work can raise scalp temperature, making it easier to activate this receptor and exacerbate hair loss. Most existing anti-hair loss products target androgen or 5α-reductase inhibition, which are unsuitable for men. Therefore, developing anti-hair loss products suitable for men is a pressing technical problem in this field. Summary of the Invention

[0004] Based on this, the purpose of the present invention is to overcome the shortcomings of the prior art and provide a hair loss prevention composition and its application containing Norway spruce leaf extract. This composition can prevent androgenetic alopecia by anti-inflammatory, antibacterial, inhibiting TRPV3 channels, inhibiting the activity of 5α-reductase, inhibiting sebaceous gland cells to secrete lipids, and achieving the purpose of oil control. Moreover, all the ingredients are plant-based, making it more suitable for men.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] In a first aspect, the present invention provides an anti-hair loss composition, the composition comprising component A and component B: component A comprises the following ingredients in parts by weight: 1-5 parts of mullein extract, 0.1-1 parts of saw palmetto extract, and 1-5 parts of Norway spruce leaf extract; component B comprises the following ingredients in parts by weight: 2-4 parts of eucommia bark extract.

[0007] Verbena officinalis extract contains verbascoside, which can inhibit TRPV3 channels, thus preventing hair loss. It also has anti-inflammatory effects by inhibiting TRPV3 channels. Saw palmetto extract can inhibit 5α-reductase activity and can be used to prevent androgenetic alopecia. Norway spruce leaf extract is rich in polysaccharides, flavonoids, and polyphenols, which can inhibit sebaceous gland cells from secreting lipids, thus controlling oil production. Inhibiting sebum secretion may also reduce inflammation to some extent. This extract also contains anti-inflammatory components such as flavonoids, thus possessing certain anti-inflammatory effects on the skin. Eucommia ulmoides bark extract can inhibit heat-induced MMP-1 gene expression, lower epidermal temperature, and has anti-inflammatory effects.

[0008] This invention has found that when the above-mentioned plant extracts are used in combination in specific amounts, they can prevent androgenetic alopecia by inhibiting inflammation, bacteria, TRPV3 channels, and 5α-reductase activity, and inhibit sebaceous gland cells to secrete lipids, thus achieving the purpose of oil control. Moreover, all of them are plant-based ingredients, making them more suitable for men.

[0009] As a preferred embodiment of the first aspect, the composition comprises the following components in parts by weight: the composition comprises component A and component B: component A comprises the following components in parts by weight: 1-4 parts of mullein extract, 0.1-0.8 parts of saw palmetto extract, and 1-4 parts of Norway spruce leaf extract; component B comprises the following components in parts by weight: 2-3 parts of Eucommia ulmoides bark extract.

[0010] In a preferred embodiment of the first aspect, the composition comprises component A and component B: component A comprises the following ingredients in parts by weight: 3 parts of mustachys extract, 0.1 parts of saw palmetto extract, and 2 parts of Norway spruce leaf extract; component B comprises the following ingredients in parts by weight: 2.5 parts of Eucommia ulmoides bark extract.

[0011] This invention has found that the mass fractions of mustachys extract, saw palmetto extract, Norway spruce leaf extract, and Eucommia bark extract in the anti-hair loss composition can affect not only the TNF-α inhibition rate and Malassezia antibacterial effect of the composition, thus affecting its anti-inflammatory, antibacterial, and antipruritic effects; but also its oil-controlling effect and its ability to inhibit 5α-reductase, thereby affecting its oil-controlling and anti-hair loss effects. When further selecting 1-5 parts of mustachys extract, 0.1-1 parts of saw palmetto extract, 1-5 parts of Norway spruce leaf extract, and 2-4 parts of eucommia bark extract, especially 1-4 parts of mustachys extract, 0.1-0.8 parts of saw palmetto extract, 1-4 parts of Norway spruce leaf extract, and 2-3 parts of eucommia bark extract, the resulting anti-hair loss composition exhibits superior oil control, anti-hair loss, anti-inflammatory, antibacterial, and antipruritic effects. When 3 parts of mustachys extract, 0.1 parts of saw palmetto extract, 2 parts of Norway spruce leaf extract, and 2.5 parts of eucommia bark extract are used, the overall comprehensive effect of the composition can be maximized.

[0012] In a preferred embodiment of the first aspect, the mass ratio of component A to component B in the composition is: component A: component B = (1.52-3.5):1. For example, it can be, but is not limited to, 1.52:1, 1.5:1, 2:1, 2.5:1, 3:1, or 3.5:1.

[0013] In a preferred embodiment of the first aspect, the mass ratio of component A to component B in the composition is: component A: component B = (1.7-2.9): 1.

[0014] In a preferred embodiment of the first aspect, the mass ratio of component A to component B in the composition is: component A: component B = 2.04:1.

[0015] This invention has found that the mass ratio of the sum of the extracts of *Mullena florida*, *Saw Palmetto*, and *Norwegian spruce* leaves to the extract of *Eucommia ulmoides* bark in the anti-hair loss composition can, to a certain extent, affect the composition's oil-controlling, anti-hair loss, anti-inflammatory, antibacterial, and antipruritic effects. Further selection of the mass ratio of (*Mullena florida* extract + *Saw Palmetto* extract + *Norwegian spruce* leaf extract) to *Eucommia ulmoides* bark extract to (1.52-3.5):1), especially (1.7-2.9):1, can enhance the composition's oil-controlling, anti-hair loss, anti-inflammatory, antibacterial, and antipruritic effects. When the mass ratio of (*Mullena florida* extract + *Saw Palmetto* extract + *Norwegian spruce* leaf extract) to *Eucommia ulmoides* bark extract is 2.04:1, the overall comprehensive effect of the composition can be maximized.

[0016] In a preferred embodiment of the first aspect, the mass ratio of the sum of the mass of the mustachys extract, the mass of the saw palmetto extract, and the mass of the Norway spruce leaf extract in composition A is: mustachys extract : (saw palmetto extract + Norway spruce leaf extract) = 1 : (1.13-2.3). For example, it can be, but is not limited to, 1:1.13, 1:1.18, 1:2, 1:2.2, or 1:2.3.

[0017] As a preferred embodiment of the first aspect, in composition A, the mass ratio of the sum of the mass of mustachys extract, the mass of saw palmetto extract, and the mass of Norway spruce leaf extract is: mustachys extract : (saw palmetto extract + Norway spruce leaf extract) = 1 : (1.17-2.1).

[0018] In a preferred embodiment of the first aspect, the mass ratio of the sum of the mass of mustachys extract, the mass of saw palmetto extract, and the mass of Norway spruce leaf extract in composition A is: mustachys extract : (saw palmetto extract + Norway spruce leaf extract) = 1 : 1.13.

[0019] This invention has found that the mass ratio of mullein extract to the sum of the masses of saw palmetto extract and Norway spruce leaf extract in a men's anti-hair loss composition can significantly affect the composition's anti-inflammatory, antibacterial, antipruritic, and oil-controlling effects. Further optimization of the mass ratio of mullein extract to (saw palmetto extract + Norway spruce leaf extract) to 1:(1.13-2.3), especially to 1:(1.17-2.1), enhances the composition's anti-inflammatory, antibacterial, antipruritic, and oil-controlling effects. A mass ratio of mullein extract to (saw palmetto extract + Norway spruce leaf extract) of 1:1.13 maximizes the overall effectiveness of the composition.

[0020] As a preferred embodiment of the first aspect, the method for preparing the mustachys extract includes the following steps: dried leaves of mustachys are dried, pulverized, and added to an aqueous solution of a eutectic solvent for ultrasonic extraction. The product is collected, centrifuged, and the supernatant is taken to obtain the mustachys extract; wherein the eutectic solvent is composed of butanediol and malic acid.

[0021] As a preferred embodiment of the first aspect, the method for preparing the mullein extract satisfies at least one of the following:

[0022] (a) The molar ratio of butanediol and malic acid is: butanediol: malic acid = 1:(1~4);

[0023] (b) The mass concentration of the eutectic solvent is 20-50%;

[0024] (c) The temperature of the ultrasonic extraction is 50-60℃;

[0025] (d) The power of the ultrasonic extraction is 60-70W;

[0026] (e) The ultrasonic extraction time is 30-60 min;

[0027] (f) The material-to-liquid ratio for ultrasonic extraction is 1 g:(20-30) mL.

[0028] This invention has found that different extraction methods affect the mullein extract and its synergistic effect with other components, thus affecting the anti-inflammatory, antibacterial, antipruritic, oil-controlling, and anti-hair loss effects of the composition, thereby affecting the overall comprehensive effect of the composition.

[0029] In a second aspect, the present invention provides the use of the anti-hair loss composition as described in the first aspect in the preparation of anti-hair loss products.

[0030] Thirdly, the present invention provides an oil-controlling and hair-preventing product, the oil-controlling and hair-preventing product comprising the male hair-preventing composition described in the first aspect.

[0031] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0032] This invention discovers that mullein extract, saw palmetto extract, Norway spruce leaf extract, and Eucommia bark extract have a synergistic effect from numerous plant extracts. The combination formed by these extracts through specific formulation can prevent androgenetic alopecia by inhibiting inflammation, bacteria, TRPV3 channel inhibition, and 5α-reductase activity, and inhibiting sebaceous gland cell lipid secretion, thereby achieving the purposes of anti-inflammatory, antibacterial, antipruritic, oil control, and hair loss prevention. Furthermore, all the ingredients are plant-based, making it more suitable for men. Detailed Implementation

[0033] To better illustrate the purpose, technical solution, and advantages of this invention, the invention will be further described below with reference to specific embodiments. The purpose is to provide a detailed understanding of the invention, not to limit it. All other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention. Unless otherwise specified, the experimental reagents and instruments designed in the embodiments and comparative examples of this invention are commonly used reagents and instruments, all of which are commercially available. Unless otherwise specified, the experimental methods used in the embodiments and comparative examples are conventional methods; and unless otherwise specified, the raw materials used in parallel experiments are from the same batch. Further explanation of the raw materials used in this invention:

[0034] Saw palmetto extract was purchased from Shaanxi Hanqing Biotechnology Co., Ltd.; product name: saw palmetto extract.

[0035] The Norway spruce leaf extract was purchased from Norture Lights Cell Science Inc., Canada. Product name: Norway spruce (PICEA EXCELSA) leaf extract.

[0036] Rehmannia glutinosa extract was purchased from Shaanxi Lvshengyuan Biological Products Manufacturing Co., Ltd., product name: Rehmannia glutinosa extract;

[0037] The Platycladus orientalis leaf extract was purchased from Shanxi Yirun Biotechnology Co., Ltd., product name: Platycladus orientalis leaf extract;

[0038] The Tremella fuciformis extract was purchased from Hubei Xinyuhong Biomedical Technology Co., Ltd., product name: Tremella fuciformis extract;

[0039] The Eucommia ulmoides extract was purchased from Shaanxi Guben Biotechnology Co., Ltd., product name: Eucommia ulmoides extract.

[0040] Preparation of Eucommia bark extract: Take commercially available Eucommia bark, dry and pulverize it, add 5 times the amount of 50% ethanol solution, stir at 50℃ for 24 hours, filter, and concentrate at 60℃ to half the original volume to obtain Eucommia bark extract.

[0041] The preparation method of mullein extract-1 is as follows:

[0042] (1) Prepare a eutectic solvent:

[0043] Take a certain amount of eutectic solvent composed of butanediol and malic acid in a molar ratio of 1:2, and dilute it with deionized water to prepare an aqueous solution of eutectic solvent with a mass concentration of 20%.

[0044] (2) Preparation of Verbena officinalis extract:

[0045] Collect dried leaves of mustachys, grind and crush them, and pass them through a 40-mesh sieve. Take 1g and add it to the aqueous solution of the above-mentioned eutectic solvent at a material-to-liquid ratio of 1g:20mL. Use ultrasonic extraction method with an ultrasonic temperature of 50℃, a power of 60W, and an extraction time of 30min. Collect the product, centrifuge, and take the supernatant to obtain the mustachys extract-1.

[0046] The preparation method of mullein extract-2 is as follows:

[0047] (1) Prepare a eutectic solvent:

[0048] Take a certain amount of eutectic solvent composed of butanediol and malic acid in a molar ratio of 1:4, and dilute it with deionized water to prepare an aqueous solution of eutectic solvent with a mass concentration of 30%.

[0049] (2) Preparation of Verbena officinalis extract:

[0050] Collect dried leaves of mustachys, grind and crush them, and pass them through a 50-mesh sieve. Take 1g and add it to the aqueous solution of the above-mentioned eutectic solvent at a material-to-liquid ratio of 1g:25mL. Use ultrasonic extraction method with an ultrasonic temperature of 55℃, a power of 65W, and an extraction time of 40min. Collect the product, centrifuge, and take the supernatant to obtain the mustachys extract-2.

[0051] The preparation method of mullein extract-3 is as follows:

[0052] (1) Prepare a eutectic solvent:

[0053] Take a certain amount of eutectic solvent composed of butanediol and malic acid in a molar ratio of 1:1, and dilute it with deionized water to prepare an aqueous solution of eutectic solvent with a mass concentration of 50%.

[0054] (2) Preparation of Verbena officinalis extract:

[0055] Collect dried leaves of mustachys, grind and crush them, and pass them through a 70-mesh sieve. Take 1g and add it to the aqueous solution of the above-mentioned eutectic solvent at a material-to-liquid ratio of 1g:30mL. Use ultrasonic extraction method with an ultrasonic temperature of 60℃, a power of 70W, and an extraction time of 60min. Collect the product, centrifuge, and take the supernatant to obtain the mustachys extract-3.

[0056] The preparation method of mullein extract-4 is as follows:

[0057] Dried leaves of mustaphyllum were collected, cleaned, and dried in a 50°C oven until constant weight. After grinding and pulverizing, the powder was passed through an 80-mesh sieve. 1 g of the powder was added to a 75% ethanol aqueous solution at a material-to-liquid ratio of 1 g:30 mL. Ultrasonic extraction was performed at 50°C, 160 W, and 20 min. The product was collected, centrifuged, and the supernatant was collected to obtain mustaphyllum extract-4.

[0058] The preparation method of mullein extract-5 is as follows:

[0059] Dried leaves of *Verbena officinalis* were collected, washed, and dried in a 50°C oven until constant weight. The dried leaves were then ground and passed through an 80-mesh sieve to obtain *Verbena officinalis* leaf powder. 1g of the powder was added to 20mL of 70% ethanol and refluxed at 80°C for 1 hour to obtain *Verbena officinalis* extract-5.

[0060] The preparation method of the male anti-hair loss composition suitable for sensitive skin of the present invention is as follows: weigh the components according to the weight parts shown in Table 1, add water to 20g, stir and mix evenly, and sonicate for 10 minutes to obtain the male anti-hair loss composition.

[0061] Examples and Comparative Examples

[0062] This application provides a hair loss prevention composition suitable for men, the components and their weight parts of the composition are shown in Table 1:

[0063] Table 1:

[0064]

[0065]

[0066] Example of effect

[0067] 1. Anti-inflammatory ability of the composition:

[0068] Plating: Logarithmically growing RAW264.7 cells were seeded into 24-well plates at a cell density of 2 × 10⁻⁶ cells / well. 5 Cells / mL, 0.5mL per well, after plating, mark the top cover of the cell plate and incubate in a CO2 incubator for 24h;

[0069] Sample processing: After cell incubation for 24 h, the supernatant was aspirated, and 0.5 mL of the following samples were added respectively: ① Blank control group: DMEM medium; ② LPS inflammation model group: LPS (1 μg / mL); ③ Positive control group: 0.001% dexamethasone + LPS (1 μg / mL); ④ Experimental group: 2% of the example or comparative composition + LPS (1 μg / mL);

[0070] Detection of inflammatory factors in cell supernatant: After 24 hours of cell treatment, the supernatant was pipetted into centrifuge tubes and centrifuged at 1000 rpm for 5 minutes. The precipitate was discarded, and the supernatant was collected. The changes in inflammatory factors in the supernatant were detected using the Xinbosheng TNF-α inflammatory factor detection kit. The inhibition rate of TNF-α was calculated as follows: TNF-α inhibition rate % = (TNF-α level in model group - TNF-α level in experimental group) / TNF-α level in model group × 100%. A higher TNF-α inhibition rate % indicates stronger anti-inflammatory ability. The calculation results are shown in Table 2.

[0071] 2. Inhibitory effect of the composition on 5α-reductase

[0072] Sample tubes: Add 1 mL of sample solution, 1 mL of enzyme solution, 1 mL of NADPH solution, and 1 mL of testosterone solution to a test tube, gently shake well, and pipette 200 μL into a 96-well ELISA plate. Perform three replicates for each sample. Place the plate in an ELISA reader and measure the absorbance at 340 nm. This is the first measurement value for sample A (0). After incubating at 37°C for 20 minutes, place the plate in an ELISA reader and measure the absorbance at 340 nm. This is the second measurement value for sample A (20).

[0073] Enzyme tubes: Add 1 mL of PBS solution, 1 mL of enzyme solution, 1 mL of NADPH solution, and 1 mL of testosterone solution to a test tube, gently shake well, and pipette 200 μL into a 96-well microplate. Perform three replicates for each sample. Place the plate in a microplate reader and measure the absorbance at 340 nm. This first measurement value is A. 酶0 After incubating the sample at 37°C for 20 minutes, it was placed in an ELISA reader for detection. The absorbance was measured at 340 nm, which is the second measurement value A. 酶20 The inhibition rate of the enzyme was calculated according to the following formula, and the results are shown in Table 2.

[0074]

[0075] The higher the 5α-reductase inhibition rate, the stronger the hair loss prevention ability.

[0076] Table 2:

[0077] Group TNF-α inhibition rate % Group 5α-reductase inhibition rate % Positive control group 45.7％ Positive control group 78.3％ Example 1 31.0％ Example 1 38.8％ Example 2 33.2％ Example 2 44.1％ Example 3 33.6％ Example 3 40.5％ Example 4 33.1％ Example 4 43.3％ Example 5 37.3％ Example 5 46.5％ Example 6 35.2％ Example 6 47.3％ Example 7 39.1％ Example 7 49.4％ Example 8 37.7％ Example 8 49.7％ Example 9 44.2％ Example 9 56.3％ Example 10 41.1％ Example 10 52.5％ Example 11 41.7％ Example 11 51.1％ Example 12 40.8％ Example 12 51.3％ Comparative Example 1 26.2％ Comparative Example 1 25.8％ Comparative Example 2 30.1％ Comparative Example 2 36.3％ Comparative Example 3 22.5％ Comparative Example 3 30.5％ Comparative Example 4 22.7％ Comparative Example 4 14.8％ Comparative Example 5 20.1％ Comparative Example 5 27.7％ Comparative Example 6 20.4％ Comparative Example 6 27.9％ Comparative Example 7 25.1％ Comparative Example 7 26.8％ Comparative Example 8 24.4％ Comparative Example 8 16.8％ Comparative Example 9 22.9％ Comparative Example 9 25.9％ Comparative Example 10 22.8％ Comparative Example 10 25.3％

[0078] As can be seen from Table 2, the anti-hair loss composition provided by the present invention can significantly improve the TNF-α inhibition rate and 5α-reductase inhibition rate, which indicates that the composition of the present invention has strong anti-inflammatory and androgen-reducing effects, thus achieving the purpose of preventing hair loss.

[0079] As can be seen from Examples 1-8 and Comparative Examples 1-2, the weight parts and mass ratios of the components in the anti-hair loss composition affect the anti-inflammatory and anti-hair loss efficacy of the composition.

[0080] Examples 1-5 show that the mass ratio of the components in the composition affects the anti-inflammatory and anti-hair loss abilities of the composition. When the mass ratio of (Mullein extract + Saw Palmetto extract + Norway Spruce leaf extract) to Eucommia bark extract is (1.52-3.5):1, the resulting composition has strong anti-inflammatory and anti-hair loss effects. When the mass ratio is further increased to (1.7-2.9):1, the composition has even better anti-inflammatory and anti-hair loss effects. The best effect is achieved when the mass ratio is 2.04:1 (Example 5).

[0081] This invention investigated the effect of the mass ratio of the sum of the masses of mustachys extract, saw palmetto extract, and Norway spruce leaf extract in the composition on the inhibition of TNF-α through Examples 6-8. Examples 6-8 show that the mass ratio of the sum of the masses of mustachys extract, saw palmetto extract, and Norway spruce leaf extract affects the composition's inhibitory effect on TNF-α, thus affecting its anti-inflammatory efficacy. When the mass ratio of mustachys extract to (saw palmetto extract + Norway spruce leaf extract) is 1:(1.13-2.3), the composition's ability to inhibit TNF-α is improved, especially at 1:(1.17-2.1), where the composition shows even better results. The optimal effect is achieved when the mass ratio is 1:1.13 (Example 7).

[0082] This invention investigated the effect of different extraction methods of mullein extract on the ability of the composition to inhibit TNF-α through Examples 9-12. Examples 9-12 show that the extraction method of mullein extract has a certain impact on the TNF-α inhibition effect, thus affecting the anti-inflammatory efficacy of the composition. The composition obtained using mullein extract-2 showed the best anti-inflammatory effect (Example 9).

[0083] The TNF-α inhibition rate of Comparative Examples 1-2 was 26-30%, and the 5α-reductase inhibition rate was 55-60%. This is because the mass ratio of each component in the compositions of Comparative Examples 1-2 was not within the range of the present invention, which affected the synergistic effect between the components and reduced the efficacy of the composition.

[0084] The TNF-α inhibition rate of Comparative Examples 3-6 was 20-22%, and the 5α-reductase inhibition rate was 40-60%. This is because the absence of one of the components in this invention in the compositions of Comparative Examples 3-6 affected the synergistic effect between the components and reduced the efficacy of the compositions.

[0085] The TNF-α inhibition rate of Comparative Examples 7-10 was 22-25%, and the 5α-reductase inhibition rate was 40-56%. This is because the compositions of Comparative Examples 7-10 replaced one of the components in the composition of the present invention with other plant extracts, which did not produce a synergistic effect with the specific components selected in the present invention, and also affected the synergistic effect between other components of the present invention, thus reducing the efficacy of the composition.

[0086] 3. Antibacterial effect test of the composition against Malassezia

[0087] Malassezia (ATCC 44344) is lipophilic, and its colonies float on the liquid surface. A direct method was used to obtain data: a single colony of Malassezia (ATCC 44344) was incubated in 3 mL of Malassezia medium (SDA agar + 1% Tween 40 + 0.6% ox bile salt + 0.001% chloramphenicol) at 37°C and 200 rpm for 48 h. The cultured Malassezia was then diluted to 10... 5 cfu / ml. The compositions from the examples and comparative examples were diluted to concentrations of 0.1%, 0.2%, 0.5%, 1%, 2%, and 5%, and 100 μL of each concentration was placed in a 96-well plate, with three replicates for each concentration. 100 μL of Malassezia bacterial suspension was added to each well, and the plate was observed for clarity. Clarity indicated an antibacterial effect. The test results are shown in Table 3 below:

[0088] Table 3:

[0089]

[0090]

[0091] As can be seen from Table 3, the anti-hair loss composition provided by the present invention can significantly inhibit Malassezia, which indicates that the composition of the present invention has strong antibacterial and antipruritic effects.

[0092] As can be seen from Examples 1-8 and Comparative Examples 1-2, the weight parts and mass ratios of the components in the anti-hair loss composition affect the antibacterial and antipruritic efficacy of the composition.

[0093] Examples 1-5 show that the mass ratio of the components in the composition affects the antibacterial ability of the composition. When the mass ratio of (Mullein extract + Saw Palmetto extract + Norway Spruce leaf extract) to Eucommia bark extract is (1.52-3.5):1, the resulting composition has a strong antibacterial and antipruritic effect. When the mass ratio is further increased to (1.7-2.9):1, the antibacterial and antipruritic effect of the resulting composition is even better. When the mass ratio is 2.04:1 (Example 5), the effect is the best.

[0094] This invention investigated the effect of the mass ratio of the sum of the masses of mustachys extract, saw palmetto extract, and Norway spruce leaf extract on the antibacterial effect of the composition through Examples 6-8. Examples 6-8 show that the mass ratio of the sum of the masses of mustachys extract, saw palmetto extract, and Norway spruce leaf extract affects the antibacterial and antipruritic effects of the composition. When the mass ratio of mustachys extract to (saw palmetto extract + Norway spruce leaf extract) is 1:(1.13-2.3), the antibacterial and antipruritic ability of the composition is improved, especially at 1:(1.17-2.1), the effect of the resulting composition is even better. The optimal effect is achieved when the mass ratio is 1:1.13 (Example 7).

[0095] This invention investigated the effect of different extraction methods of mullein extract on the antibacterial activity of the composition through Examples 9-12. Examples 9-12 show that the extraction method of mullein extract has a certain impact on the antibacterial effect, thus affecting the antibacterial and antipruritic efficacy of the composition. The composition obtained using mullein extract-2 showed the best antibacterial and antipruritic effect (Example 9).

[0096] The antibacterial effect of Comparative Examples 1-2 was poor. This is because the mass ratio of each component in the composition of Comparative Examples 1-2 was not within the range of the present invention, which affected the synergistic effect between the components and reduced the efficacy of the composition.

[0097] The antibacterial effects of Comparative Examples 3-6 were poor because the absence of one of the components in this invention in the compositions of Comparative Examples 3-6 affected the synergistic effect between the components and reduced the efficacy of the compositions.

[0098] The antibacterial effect of Comparative Examples 7-10 was poor. This is because the compositions of Comparative Examples 7-10 replaced one of the components in the composition of the present invention with other plant extracts, which did not produce a synergistic effect with the specific components selected in the present invention, and also affected the synergistic effect between other components of the present invention, thus reducing the efficacy of the composition.

[0099] Application examples

[0100] The present invention provides a men's anti-hair loss product in application examples and comparative application examples. The components (mass percentage) of the anti-hair loss product are shown in Table 4. In application examples 1-12, the men's anti-hair loss compositions used are the same as those prepared in examples 1-12. In application examples 13-14, the compositions prepared in example 9 are used. In comparative application examples 1-10, the men's anti-hair loss compositions used are the same as those prepared in comparative examples 1-10. A blank application example is also provided, the only difference between the blank application example and application example 1 being that no men's anti-hair loss composition is added.

[0101] Table 4:

[0102]

[0103]

[0104] This embodiment provides a method for preparing shampoo, the method comprising the following steps:

[0105] (1) Add polyquaternium-10 to deionized water and stir to disperse. After the dispersion is uniform, add other materials of phase A, heat to 85°C, stir to dissolve evenly, and then keep warm for later use to obtain pre-phase A.

[0106] (2) Add each component of phase B at 80-85℃, stir thoroughly to dissolve evenly until transparent, and keep warm to defoam;

[0107] (3) Cool down to below 45℃, add the C phase raw materials one by one, and stir evenly;

[0108] (4) Continue to add phase D raw materials and mix evenly to obtain men's anti-hair loss shampoo.

[0109] Test Example 1: Oil Control Effect Test of Anti-Hair Loss Products

[0110] Several male volunteers aged 18-45 were selected. Subjects sat quietly for 30 minutes in a constant temperature and humidity room (21±1℃, 50±10%). Initial sebum content was measured using a Meibometer MB560 scalp sebum analyzer, with the test site being the top of the scalp. Subjects were randomly divided into 25 groups of 5 people each, and randomly assigned to use the anti-hair loss product provided by this invention. Testing was conducted at 8 and 12 weeks, and the improvement in sebum content for each volunteer at 8 and 12 weeks was analyzed. Improvement was expressed as the sebum reduction rate, calculated as follows:

[0111] The reduction rate relative to the initial value % = |(X 使用后 -X 使用前 ) / X 使用前 ×100% |; where, the larger the percentage reduction relative to the initial value, the stronger the anti-hair loss ability. The calculated data are shown in Table 5;

[0112] Table 5:

[0113]

[0114]

[0115] As can be seen from Table 5, the anti-hair loss product provided by the present invention can significantly reduce oil secretion, which indicates that the composition of the present invention has a strong oil control and anti-hair loss effect.

[0116] As can be seen from Application Examples 1-8 and Comparative Application Examples 1-2, the weight parts and mass ratios of the components in the anti-hair loss composition affect the oil control and anti-hair loss efficacy of the composition.

[0117] As can be seen from Application Examples 1-5, the mass ratio of the components in the composition affects the antibacterial ability of the composition. When the mass ratio of (Mullein extract + Saw Palmetto extract + Norway Spruce leaf extract) to Eucommia bark extract is (1.52-3.5):1, the resulting composition has a strong oil-controlling and hair-preventing effect. When it is further reduced to (1.7-2.9):1, the oil-controlling and hair-preventing effect of the resulting composition is even better. When the mass ratio is 2.04:1 (Application Example 5), the effect is the best.

[0118] This invention investigated the effect of the mass ratio of the sum of the masses of mustachys extract, saw palmetto extract, and Norway spruce leaf extract in the composition on the oil-controlling and hair-preventing effects of the composition through Application Examples 6-8. It can be seen that the mass ratio of the sum of the masses of mustachys extract, saw palmetto extract, and Norway spruce leaf extract affects the oil-controlling and hair-preventing effects of the composition. When the mass ratio of mustachys extract to (saw palmetto extract + Norway spruce leaf extract) is 1:(1.13-2.3), the oil-controlling and hair-preventing ability of the composition is improved, especially when the ratio is 1:(1.17-2.1), the effect of the resulting composition is even better. The effect is optimal when the mass ratio is 1:1.13 (Application Example 7).

[0119] This invention investigated the effects of different extraction methods of *Verbena officinalis* extract on the oil-controlling and hair-preventing efficacy of the composition through Application Examples 9-12. It can be seen that the extraction method of *Verbena officinalis* extract has a certain impact on sebum secretion, thus affecting the oil-controlling and hair-preventing efficacy of the composition. The composition obtained using *Verbena officinalis* extract-2 showed the best oil-controlling and hair-preventing effect (Application Example 9).

[0120] The oil reduction rate of Comparative Examples 1-2 was 16-18%. This is because the mass ratio of each component in the composition of Comparative Examples 1-2 was not within the range of the present invention, which affected the synergistic effect between the components and reduced the efficacy of the composition.

[0121] The oil reduction rate of Comparative Examples 3-6 was 14-15%, which is because the absence of one of the components of the present invention in the compositions of Comparative Examples 3-6 affected the synergistic effect between the components and reduced the efficacy of the composition.

[0122] The oil reduction rate of Comparative Examples 7-10 was 15-16%. This is because the compositions of Comparative Examples 7-10 replaced one of the components in the composition of the present invention with other plant extracts, which did not produce a synergistic effect with the specific components selected in the present invention, and also affected the synergistic effect between other components of the present invention, thus reducing the efficacy of the composition.

[0123] Test Example 2: Test on the anti-hair loss effect of anti-hair loss products on the human body

[0124] Several male volunteers aged 18-45 with varying degrees of hair loss were selected and divided into 25 groups of 5 people each. Participants used the hair loss prevention product (either the example or control product) once daily. Hair loss was counted after 8 and 12 weeks of use, using the 60-comb test to record the number of hairs lost during each combing. The number of hairs lost before product use and after 8 and 12 weeks of use was recorded. Participants did not wash their hair for 48 ± 4 hours before each visit and did not comb their hair on the day of the visit. The hair loss reduction rate was calculated as follows: Hair loss reduction rate / % = (Number of hairs lost at week 0 - Number of hairs lost at weeks 8 / 12) / Number of hairs lost at week 0. The calculated data are shown in Table 6.

[0125] Table 6

[0126]

[0127]

[0128] As can be seen from Table 6, the hair loss prevention product provided by this invention has excellent hair loss prevention effects. Specifically, at week 12, the hair loss reduction rate was over 40%.

[0129] As can be seen from Application Examples 1-8 and Comparative Application Examples 1-2, the weight parts and mass ratios of the components in the anti-hair loss composition affect the oil control and anti-hair loss efficacy of the composition.

[0130] As can be seen from Application Examples 1-5, the mass ratio of the components in the composition affects the composition's anti-hair loss ability. When the mass ratio of (Mullein extract + Saw Palmetto extract + Norway Spruce leaf extract) to Eucommia bark extract is (1.52-3.5):1, the resulting composition has a strong anti-hair loss effect. When the mass ratio is further increased to (1.7-2.9):1, the anti-hair loss effect is even better. When the mass ratio is 2.04:1 (Application Example 5), the effect is optimal.

[0131] This invention investigated the effect of the mass ratio of mustachys extract, saw palmetto extract, and Norway spruce leaf extract on the anti-hair loss effect of the composition through Application Examples 6-8. It can be seen that the mass ratio of the total mass of mustachys extract, saw palmetto extract, and Norway spruce leaf extract affects the anti-hair loss effect of the composition. When the mass ratio of mustachys extract to (saw palmetto extract + Norway spruce leaf extract) is 1:(1.13-2.3), the anti-hair loss ability of the composition is improved, especially when the ratio is 1:(1.17-2.1), the effect of the resulting composition is even better. The optimal effect is achieved when the mass ratio is 1:1.13 (Application Example 7).

[0132] This invention investigated the effect of different extraction methods of mullein extract on the anti-hair loss ability of the composition through Application Examples 9-12. It can be seen that the extraction method of mullein extract affects the anti-hair loss efficacy of the composition. The composition obtained using mullein extract-2 showed the best anti-hair loss effect (Application Example 9).

[0133] The hair loss reduction rate of Comparative Examples 1-2 was 36-38%. This is because the mass ratio of each component in the composition of Comparative Examples 1-2 was not within the scope of this invention, which affected the synergistic effect between the components and reduced the efficacy of the composition.

[0134] The hair loss reduction rate of Comparative Examples 3-6 was 33%. This is because the absence of one of the components of the present invention in the compositions of Comparative Examples 3-6 affected the synergistic effect between the components and reduced the efficacy of the composition.

[0135] The hair loss reduction rate of Comparative Examples 7-10 was 34-36%. This is because the compositions of Comparative Examples 7-10 replaced one of the components in the composition of the present invention with other plant extracts, which did not produce a synergistic effect with the specific components selected in the present invention, and also affected the synergistic effect between other components of the present invention, thus reducing the efficacy of the composition.

[0136] Test Example 3: Anti-hair loss product usage test for male consumers:

[0137] The efficacy examples of this invention verify the suitability of the anti-hair loss product prepared in Application Example 1 for male consumers and its anti-hair loss and soothing effects, specifically including the following steps:

[0138] Thirty male subjects were selected to participate in the test. They were given the product and each subject applied an appropriate amount daily for a 28-day trial. Consumers completed a questionnaire survey on the degree of improvement in indicators such as anti-hair loss effect, soothing effect, oil control effect, and irritation. See Table 7 for details.

[0139] Table 7:

[0140]

[0141] As can be seen from Table 7, the anti-hair loss product of the present invention with added anti-hair loss composition has the effects of preventing hair loss, soothing, and relieving itching, and can allow the subjects to experience a refreshing and comfortable feeling. At the same time, the anti-hair loss product of the present invention is gentle and non-irritating, and is suitable for men to use.

[0142] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A hair loss prevention composition, characterized in that, The composition consists of component A and component B: Component A consists of the following ingredients in parts by weight: 1-5 parts of mullein extract, 0.1-1 parts of saw palmetto extract, and 1-5 parts of Norway spruce leaf extract. Component B is 2-4 parts of Eucommia ulmoides bark extract; The preparation method of the mustachys extract includes the following steps: dried mustachys leaves are dried, pulverized, and added to an aqueous solution of a eutectic solvent for ultrasonic extraction. The product is collected, centrifuged, and the supernatant is taken to obtain the mustachys extract. The eutectic solvent is composed of butanediol and malic acid. The preparation method of the mustachys extract meets the following conditions: (a) The molar ratio of butanediol to malic acid is: butanediol: malic acid = 1: (1~4). (b) The mass concentration of the eutectic solvent in the aqueous solution of the eutectic solvent is 20-50%; (c) The temperature of the ultrasonic extraction is 50-60℃; (d) The power of the ultrasonic extraction is 60-70W; (e) The ultrasonic extraction time is 30-60 min; (f) The material-to-liquid ratio for ultrasonic extraction is 1g:(20~30)mL; The preparation method of the Eucommia bark extract includes the following steps: take Eucommia bark, dry and pulverize it, add 5 times the amount of 50% ethanol solution, stir at 50°C for 24 hours, filter, and concentrate at 60°C to half the original volume to obtain Eucommia bark extract.

2. The anti-hair loss composition according to claim 1, characterized in that, The composition consists of component A and component B: component A consists of the following ingredients in parts by weight: 1-4 parts of mullein extract, 0.1-0.8 parts of saw palmetto extract, and 1-4 parts of Norway spruce leaf extract; component B consists of 2-3 parts of Eucommia ulmoides bark extract.

3. The anti-hair loss composition as described in claim 1, characterized in that, In the composition, the mass ratio of component A to component B is: component A: component B = (1.52-3.5):

1.

4. The anti-hair loss composition according to claim 3, characterized in that, In the composition, the mass ratio of component A to component B is: component A: component B = (1.7-2.9):

1.

5. The anti-hair loss composition as described in claim 1, characterized in that, In composition A, the mass ratio of the sum of the mass of mustachys extract, the mass of saw palmetto extract, and the mass of Norway spruce leaf extract is: mustachys extract : (saw palmetto extract + Norway spruce leaf extract) = 1 : (1.13-2.3).

6. The anti-hair loss composition as described in claim 5, characterized in that, In the composition A, the mass ratio of the sum of the mass of mustachys extract, the mass of saw palmetto extract, and the mass of Norway spruce leaf extract is: mustachys extract : (saw palmetto extract + Norway spruce leaf extract) = 1 : (1.17-2.1).

7. The use of the anti-hair loss composition according to any one of claims 1-6 in the preparation of anti-hair loss products.

8. An oil-controlling and hair-preventing product, characterized in that, The oil-controlling and hair-preventing product includes the hair-preventing composition as described in any one of claims 1-6.

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