Hair loss preventing and hair growing composition and application thereof

Abstract

The invention relates to an anti-hair-loss hair-growing composition, which is formed by homogenizing and compounding active components, namely a nasturtium officinale extract, a peony root extract, a ginkgo leaf extract, a scutellaria baicalensis root extract, a sophora flavescens root extract and a soybean lactobacillus fermentation product in a butanediol and / or pentanediol-water solvent system. A multi-channel synergistic anti-hair loss action mechanism of hair follicle activation-micro-ecological balance-inflammation inhibition is constructed, scalp health is improved from multiple dimensions, and the composition is suitable for improving the problems of seborrheic alopecia, pressure alopecia, scalp micro-ecological environment imbalance and the like.

Description

Technical Field

[0001] This invention belongs to the field of functional cosmetics technology, and relates to a plant compound composition with anti-hair loss and hair growth effects, and its application in hair care products. Background Technology

[0002] Hair loss is showing a significant upward trend globally, with the age of onset becoming increasingly younger. This phenomenon is closely related to the stresses of modern society, changes in lifestyle, and external environmental factors. Hair loss not only affects a patient's appearance but also easily triggers mental health problems, thereby creating a growing demand in the hair treatment market.

[0003] Hair loss prevention and hair regrowth technology has become a research hotspot in the medical and beauty industries. It mainly includes three major directions: chemical drug intervention, surgical procedures, and the application of plant active ingredients. However, each technology has obvious limitations.

[0004] In the field of chemical drugs, 5α-reductase inhibitors, represented by finasteride, are the first-line treatment for male pattern baldness by specifically blocking the androgen pathway. However, adverse reactions such as sexual dysfunction, anxiety, and abnormal breast tissue hyperplasia have been consistently reported in clinical applications, severely limiting their long-term safety. Hair transplantation surgery, as a physical intervention, faces bottlenecks such as high treatment costs and long postoperative recovery periods. While single plant extracts such as ginger and Polygonum multiflorum have certain hair growth activity, they generally have insufficient efficacy and may induce contact dermatitis.

[0005] In recent years, plant-based hair loss prevention technology has gradually shifted towards enhancing efficacy through the synergistic effects of multiple components and process optimization. For example, it uses a specific mass ratio of (1-3):2, combining arborvitae leaf extract (which inhibits 5α-reductase activity) and Polygonum multiflorum extract (which promotes hair follicle cycle transition), to prevent hair loss by inhibiting the androgen pathway; some improved formulas introduce active peptides (such as marine shellfish hydrolysates) to enhance the nutrient supply to hair follicles; or use lactobacillus fermentation products to regulate the scalp's microecological balance.

[0006] However, these technical approaches still share the following common technical shortcomings:

[0007] 1. Insufficient multi-pathway synergistic mechanism: Existing compound systems mostly focus on the mechanism of action of a single component, lacking synergistic intervention and systematic integration of key mechanisms such as improvement of hair follicle microcirculation, inhibition of inflammatory factors and activation of hair follicle stem cells, especially neglecting the cause of seborrheic alopecia caused by Malassezia imbalance;

[0008] 2. Low transdermal delivery efficiency: Plant extracts have a large molecular weight, and large molecular plant active ingredients (such as Polygonum multiflorum glycoside with a transdermal transdermal rate of <15%) are difficult to effectively penetrate the scalp barrier, resulting in insufficient effective concentration in the hair follicle microenvironment.

[0009] 3. Ingredient stability defects: The active peptides in the compound system are prone to thermal degradation and acid hydrolysis under normal storage conditions, with an activity decay rate of more than 40% within 6 months, which significantly affects the consistency of efficacy within the product's shelf life.

[0010] 4. Disconnection in microecological regulation: Although some studies have used probiotic fermentation products such as lactobacillus to regulate the balance of scalp flora, they are mostly used as single additives and have failed to establish a metabolic synergy or signal transduction linkage mechanism with the core hair growth ingredients.

[0011] The aforementioned technological bottlenecks prevent existing plant-based hair loss prevention products from achieving synergistic regulation across the entire pathway of "hair follicle activation - microecological balance - inflammation suppression," thus limiting their clinical hair regrowth efficacy. Therefore, the development of novel hair loss prevention and regrowth systems that combine multi-target action, highly efficient transdermal properties, and ingredient stability has become an urgent industry need. Summary of the Invention

[0012] The purpose of this invention is to provide a hair loss prevention and hair growth composition and its application, which achieves multiple mechanisms of action such as "hair follicle activation-microecological balance-inflammation inhibition" through a multi-pathway synergistic mechanism, in order to improve problems such as seborrheic alopecia, stress-induced alopecia and scalp microecological imbalance.

[0013] The hair loss prevention and hair growth composition of the present invention is composed of active ingredients and a solvent system:

[0014] The active ingredients include watercress ( Nasturtium officinale ) extract, peony ( Paeonia lactiflora ) root extract, Ginkgo ( Ginkgo biloba Leaf extract, Scutellaria baicalensis ( Scutellaria baicalensis ) root extract, Sophora flavescens ( Sophora flavescens ) root extract, soybean ( Glycine max ) Lactobacillus ( Lactobacillus (spp.) fermentation products;

[0015] The solvent system is an aqueous solution of butanediol and / or pentanediol;

[0016] The above components are homogenized and compounded to form a composition that inhibits hair loss and promotes hair follicle activity.

[0017] Specifically, the active ingredients contained in the anti-hair loss and hair growth composition of the present invention are as follows: 1-20% watercress extract, 1-10% peony root extract, 0.1-0.3% ginkgo leaf extract, 0.1-0.3% scutellaria root extract, 0.5-1% sophora root extract, and 0.1-0.5% Lactobacillus soyba fermentation product.

[0018] Furthermore, the solvent system in the hair loss prevention and hair growth composition of the present invention is composed of 20-30% by mass of butylene glycol and / or pentanediol and the balance water. Through the small molecule penetration technology of the butylene glycol and / or pentanediol solvent system, the transdermal permeability of the composition is increased to ≥30%.

[0019] The active components in this invention, including watercress extract, peony root extract, ginkgo leaf extract, scutellaria root extract, and sophora root extract, can all be prepared using any conventional extraction process for active components of traditional Chinese medicine, such as water extraction, water extraction with alcohol precipitation, ethanol hot reflux extraction, and other extraction methods. This invention does not have any special requirements for them.

[0020] Furthermore, and particularly, the present invention preferably employs a preparation process in which dehydrated and dried watercress is extracted by hot reflux extraction with a 50% ethanol solution, concentrated, and then freeze-dried to obtain watercress extract. The ethanol hot reflux extraction + freeze-drying process can increase the purity of watercress flavonoids in the extract to ≥15.5 wt%, ensuring the concentration of active ingredients.

[0021] Alternatively, a preferred method is to extract peony root by hot reflux extraction with ethanol, followed by filtration through macroporous adsorption resin and concentration under reduced pressure to obtain a concentrated solution, which is then freeze-dried to obtain peony root extract.

[0022] For both Ginkgo biloba leaf extract and Scutellaria baicalensis root extract, the preferred method is to extract them by heating with pure water, then concentrate under reduced pressure, decolorize by activated carbon adsorption, filter to obtain the filtrate, and freeze-dry to obtain the extract.

[0023] The preferred method for extracting Sophora flavescens root extract is hot water reflux extraction. The extract is concentrated under reduced pressure, and ethanol is added for alcohol precipitation. The supernatant is collected by centrifugation, concentrated under reduced pressure, and then freeze-dried to obtain Sophora flavescens root extract.

[0024] Furthermore, the soybean lactic acid bacteria fermentation product in this invention is a fermentation product with a solid content of ≥0.18wt% obtained by inoculating ground soybeans with lactic acid bacteria and fermenting them.

[0025] Among the various active ingredients mentioned above, watercress extract has the effects of clearing heat and detoxifying, promoting diuresis, relieving phlegm and cough, cooling blood and stopping bleeding. Its rich content of flavonoids and phenolic acids has antioxidant and anti-hair loss factor effects. Peony root extract can promote blood circulation and regulate immunity; its paeoniflorin content has good anti-inflammatory, soothing, antioxidant, and immune-regulating effects. Ginkgo leaf extract can clear the meridians and relieve pain, promote blood circulation and remove blood stasis; its rich content of flavonoids and terpene lactones has strong antioxidant effects, can improve skin microcirculation, and protect collagen and elasticity. Protein; Scutellaria baicalensis root extract has the effects of clearing heat and drying dampness, purging fire and detoxifying. Its main active ingredient, baicalin, has significant anti-inflammatory and anti-allergic effects, and can also be antibacterial and control oil; Sophora flavescens root extract has the effects of clearing heat and drying dampness, killing insects and relieving itching. The alkaloids and flavonoids it contains can have anti-inflammatory and soothing effects, relieve skin itching and discomfort, and balance the skin microecology; In the fermentation products of Lactobacillus sacchariformis, the isoflavones contained in soybeans can inhibit 5α-reductase, which helps to balance oil secretion. After further fermentation by Lactobacillus sacchariformis, it can help improve atopic dermatitis and strengthen the skin barrier.

[0026] Therefore, the anti-hair loss and hair growth composition provided by the present invention achieves the effect of improving scalp health through multiple pathways through the synergistic effect of various active components.

[0027] First, the combination of watercress extract and ginkgo leaf extract is beneficial. The flavonoids and phenolic acids in watercress can promote hair follicle growth, while the flavonoid glycosides and terpene lactones in ginkgo leaf extract can enhance the microcirculation of capillaries in the lower part of the hair follicle and improve blood supply to the scalp. The two work together to reduce free radical damage to hair follicles and alleviate oxidative stress.

[0028] Secondly, the combination of Scutellaria baicalensis root extract and Paeonia lactiflora root extract can effectively inhibit the release of inflammatory factors. Experiments have shown that this combination can reduce TNF-α levels by up to 65%, significantly alleviate perifollicular inflammatory response, and significantly improve scalp redness and inflammation caused by seborrheic dermatitis.

[0029] Furthermore, the synergistic effect of Sophora flavescens root extract and Lactobacillus sacchariformis fermentation products is as follows: Sophora flavescens alkaloids disrupt the fungal cell membrane and inhibit Malassezia colonization; while the post-biotic active ingredients contained in Lactobacillus sacchariformis fermentation products help to adjust the scalp pH to a slightly acidic environment of 5.5-6.0, inhibiting the proliferation of pathogenic bacteria, while reducing excessive sebum secretion and lowering the risk of seborrheic alopecia.

[0030] In summary, the hair loss prevention and hair growth composition of the present invention integrates hair follicle activating agents, scalp microecological regulators (lactobacillus fermentation products) and anti-inflammatory components. Through their combination, a multi-pathway synergistic mechanism for preventing hair loss is constructed, which can be used to improve scalp condition from multiple dimensions and is suitable for preventing and treating hair loss problems caused by various reasons.

[0031] The production process of the hair loss prevention and hair growth composition of the present invention is simple. By using different extraction processes to extract different raw materials in a targeted manner, the extraction efficiency is improved, the production cost is reduced, and it is suitable for large-scale production.

[0032] The hair loss prevention and hair growth composition of the present invention can be used as the main active ingredient, and by adding various conventional components, it can be used to prepare various hair care products including but not limited to hair loss prevention and hair growth shampoo, hair loss prevention conditioner / hair mask, hair loss prevention spray / serum, hair loss prevention oil / essence, etc.

[0033] The preferred amount of the hair loss prevention and hair growth composition of the present invention added to various hair care products is 0.1 to 20 wt%. Attached Figure Description

[0034] Figure 1 These are the DPPH clearance rate results of the anti-hair loss and hair growth compositions in different embodiments.

[0035] Figure 2 This describes the effect of different concentrations of anti-hair loss and hair growth composition on promoting the proliferation of HDP cells.

[0036] Figure 3 The study investigated the inhibitory effect of different concentrations of anti-hair loss and hair growth compositions on TGF-β in HDP cells.

[0037] Figure 4 These are the results of hair growth experiments on different groups of mice.

[0038] Figure 5 This is a comparison of hair growth and hair follicle improvement in human trials of anti-hair loss and hair growth compositions. Implementation

[0039] The specific embodiments of the present invention will be further described in detail below with reference to the accompanying drawings and examples. The following examples are only used to more clearly illustrate the technical solutions of the present invention, so that those skilled in the art can better understand and utilize the present invention, and are not intended to limit the scope of protection of the present invention.

[0040] Unless otherwise specified, the production processes, experimental methods, or testing methods involved in the embodiments of this invention are all conventional methods in the prior art, and their names and / or abbreviations are all conventional names in the field, which are very clear and distinct in the relevant application areas. Those skilled in the art can understand the conventional process steps based on the names and apply the corresponding equipment, and implement them according to conventional conditions or the conditions recommended by the manufacturer.

[0041] The various instruments, equipment, raw materials or reagents used in the embodiments of this invention are not subject to any special restrictions on their source. They are all conventional products that can be purchased through regular commercial channels and can be prepared according to conventional methods known to those skilled in the art.

[0042] The anti-hair loss and hair growth composition in the following embodiments of the present invention is prepared using watercress extract, peony root extract, ginkgo leaf extract, scutellaria root extract, sophora root extract, and fermentation product of Lactobacillus soyba as active ingredients, wherein each active ingredient is prepared according to the following method.

[0043] Preparation method of watercress extract:

[0044] 1) Select the whole watercress plant, screen to remove impurities and rotten roots, and wash it clean;

[0045] 2) After cleaning the watercress, put it into a drying oven to dehydrate and dry it, then crush it into powder using a grinder;

[0046] 3) Use 50% ethanol to extract the crushed watercress powder by hot reflux, collect the extract, and concentrate it under reduced pressure to obtain a thick paste-like extract.

[0047] 4) The above extract was dried in a freeze dryer at 10-15 Pa and -50°C for 24-36 hours to remove residual moisture, and then pulverized to obtain watercress extract.

[0048] Preparation method of peony root extract:

[0049] 1) Peel and slice the peony root, then sun-dry it;

[0050] 2) Use ethanol to perform hot reflux extraction on dried peony roots;

[0051] 3) After the extract flows through the macroporous adsorption resin, it is concentrated under reduced pressure to obtain the concentrate;

[0052] 4) Freeze-dry the concentrate to obtain peony root extract.

[0053] Ginkgo biloba leaf extract and Scutellaria baicalensis root extract were prepared using the same method:

[0054] 1) After coarsely crushing the raw materials, add water and heat to extract them;

[0055] 2) The extract is concentrated under reduced pressure, decolorized by activated carbon adsorption, and then filtered to obtain the concentrated solution;

[0056] 3) Freeze-dry the concentrate to obtain the extract.

[0057] Preparation method of Sophora flavescens root extract:

[0058] 1) Take dried Sophora flavescens medicinal material, add water and perform hot reflux extraction;

[0059] 2) Collect the extract and concentrate it under reduced pressure to obtain a concentrated solution;

[0060] 3) Add ethanol until the final alcohol content reaches 70%, let stand at low temperature for 12 hours, and collect the supernatant by centrifugation;

[0061] 4) The supernatant was distilled under reduced pressure and the concentrated liquid was recovered and freeze-dried to obtain Sophora flavescens extract.

[0062] Preparation method of fermentation products of Lactobacillus soybaby:

[0063] 1) Soak soybeans in water, grind them, heat them to sterilize, and then inoculate them with lactic acid bacteria for fermentation;

[0064] 2) After sterilization and disinfection of the fermentation broth, the solid content is adjusted to ≥0.18% to prepare the fermentation product of Lactobacillus soybaby.

[0065] Using the raw materials prepared above as active components, the following active components are mixed evenly according to their mass percentages: watercress extract 1-20%, peony root extract 1-10%, ginkgo leaf extract 0.1-0.3%, scutellaria root extract 0.1-0.3%, sophora root extract 0.5-1%, and Lactobacillus fermentation product of soybean 0.1-0.5%. Then, 20-30% butylene glycol and / or pentylene glycol are added, and water is added to make up to 100%. After homogenization, the hair loss prevention and hair growth composition of the present invention is formed.

[0066] The hair loss prevention and hair growth composition prepared in this invention can be integrated into at least the following three categories of hair care products through functional compounding, covering the entire process of scalp care.

[0067] 1. Wash-off products that primarily focus on cleansing and conditioning:

[0068] 1) Anti-hair loss and hair growth shampoo: Add 10-20 wt% of the anti-hair loss and hair growth composition of the present invention to anionic surfactant system (such as sodium lauryl ether sulfate), and combine with conditioning agents (such as 0.05-2 wt% cationic polymer) to enhance the shampoo's retention;

[0069] 2) Anti-hair loss conditioner / hair mask: The anti-hair loss and hair growth composition of the present invention is incorporated into an emulsion system (such as cetearyl alcohol + silicone oil), and adsorbed onto the hair shaft and hair follicles by cationic charge.

[0070] 2. No-rinse products:

[0071] 1) Anti-hair loss spray / serum: Using ethanol / polyol as a solvent carrier, it is sprayed directly onto the scalp. The amount of the anti-hair loss and hair growth composition of this invention added is 5-15 wt%.

[0072] 2) Anti-hair loss oil / essential oil: Mixed with plant base oil (castor oil, sesame oil, etc.), the proportion of the anti-hair loss and hair growth composition of this invention is 8-12 wt%, and the penetration is promoted by massage.

[0073] 3. Special dosage forms such as liposome encapsulation systems:

[0074] The hair loss prevention and hair growth composition of the present invention is encapsulated in a phospholipid bilayer (such as lecithin) to improve bioavailability, and is suitable for high-concentration essences with a hair loss prevention and hair growth composition content of 15-20 wt%. Example

[0075] Example 1

[0076] The formula for the anti-hair loss and hair growth composition is as follows: watercress extract 10wt%, peony root extract 6wt%, ginkgo leaf extract 0.2wt%, scutellaria root extract 0.2wt%, sophora root extract 0.8wt%, Lactobacillus soyba ferment product 0.3wt%, butylene glycol 25wt%, and purified water to make up to 100wt%.

[0077] The hair loss prevention and hair growth composition of this embodiment takes into account both hair follicle activation and microecological balance, and is suitable for use in preparing basic balancing hair loss prevention essence, which can be directly sprayed onto the scalp.

[0078] Example 2

[0079] The formula for the anti-hair loss and hair growth composition is as follows: watercress extract 20wt%, peony root extract 1wt%, ginkgo leaf extract 0.3wt%, scutellaria root extract 0.1wt%, sophora root extract 0.5wt%, Lactobacillus soyba ferment product 0.1wt%, butylene glycol 20wt%, and purified water to make up to 100wt%.

[0080] The hair loss prevention and hair growth composition of this embodiment is mainly used to strengthen hair follicle growth factors, and is suitable for preparing hair loss prevention films for high-concentration local care, and is applicable to androgenetic alopecia.

[0081] Example 3

[0082] The formula for the anti-hair loss and hair growth composition is as follows: watercress extract 1wt%, peony root extract 10wt%, ginkgo leaf extract 0.1wt%, scutellaria root extract 0.3wt%, sophora root extract 1wt%, Lactobacillus soyba ferment product 0.5wt%, butylene glycol 30wt%, and purified water to make up to 100wt%.

[0083] The hair loss prevention and hair growth composition of this embodiment is characterized by inhibiting TNF-α and enhancing the antifungal effect of Sophora flavescens, making it suitable for preparing anti-inflammatory essences that can be directly applied to the scalp for seborrheic dermatitis.

[0084] Example 4

[0085] The formula for the anti-hair loss and hair growth composition is as follows: watercress extract 5wt%, peony root extract 3wt%, ginkgo leaf extract 0.15wt%, scutellaria root extract 0.15wt%, sophora root extract 0.6wt%, Lactobacillus soyba ferment product 0.5wt%, butylene glycol 22wt%, and purified water to make up to 100wt%.

[0086] The hair loss prevention and hair growth composition of this embodiment is suitable for preparing oil-control shampoo by compounding anionic surfactant system, maintaining the pH of the scalp environment at 5.5, and inhibiting Malassezia colonization.

[0087] Example 5

[0088] The formula for the anti-hair loss and hair growth composition is as follows: watercress extract 1wt%, peony root extract 1wt%, ginkgo leaf extract 0.1wt%, scutellaria root extract 0.1wt%, sophora root extract 0.5wt%, Lactobacillus soyba ferment product 0.1wt%, butylene glycol 20wt%, and purified water to make up to 100wt%.

[0089] The hair loss prevention and hair growth composition of this embodiment balances efficacy and cost control, and is suitable for preparing basic care type hair loss prevention conditioner.

[0090] Example 6

[0091] The formula for the anti-hair loss and hair growth composition is as follows: watercress extract 15wt%, peony root extract 8wt%, ginkgo leaf extract 0.25wt%, scutellaria root extract 0.25wt%, sophora root extract 0.7wt%, Lactobacillus soyba ferment product 0.3wt%, butylene glycol 28wt%, azone, a penetration enhancer 2wt%, and purified water to make up to 100wt.

[0092] The hair loss prevention and hair growth composition of this embodiment is suitable for preparing nanoliposome essence. The addition of azone enhances transdermal efficiency and is suitable for stubborn hair loss.

[0093] Example 7

[0094] Add 1–1.8 g of hydroxypropyl guar propyltrimethylammonium chloride to a reaction vessel containing 700 mL of purified water. Stir thoroughly until the mixture becomes transparent. Heat to 75–80 °C, and while stirring, add 1–1.8 g of sodium C14-16 olefin sulfonate, 30–40 g of lauryl hydroxysulfonate betaine, 15–20 g of CMMEA (cocamidomethyl MEA, glycerol), 40–50 g of sodium methyl cocoyl taurate, and 30–40 g of other ingredients sequentially. 0.5–0.8 g lauryl glucoside, 2–5 g disodium EDTA, 1–3 g methyl glucoside (PEG-120), and 1–3 g sodium benzoate were completely dissolved and kept warm for 10–15 minutes. The temperature was then lowered to 45°C, and 2–5 g phenoxyethanol, 10–100 g of the anti-hair loss and hair growth composition prepared in Example 4, and 2.5–5 g daily fragrance were added. Water was added to make up to 1000 mL, and the mixture was stirred evenly to prepare the anti-hair loss and hair growth shampoo.

[0095] Example 8

[0096] Add 0.1–0.2 g of allantoin to a reaction vessel containing 20 mL of purified water. After thorough stirring and dispersion, add 0.5–1.0 g of p-hydroxyacetophenone and 0.5–1.0 g of 1,2-hexanediol. Stir and heat to 85°C, keep warm for 30 min and stir until completely dissolved. After cooling to 45°C, add 10–20 g of the anti-hair loss and hair growth composition prepared in Example 1, 0.15–0.35 g of panthenol, 0.05–0.10 g of dextran, and 0.1–0.2 g of PEG-40 hydrogenated castor oil in sequence. After stirring evenly, finally adjust the pH value to 4.5–5.5 to prepare the anti-hair loss essence.

[0097] Example 9

[0098] 1,1-Diphenyl-2-trinitrophenylhydrazine (DPPH) is a stable organic free radical whose ethanol solution is purple, exhibiting a maximum absorption peak at 517 nm. When an antioxidant that can donate hydrogen atoms or electrons is present in the system, the DPPH radical is reduced to a colorless 1,1-diphenyl-2-trinitrophenylhydrazine derivative, causing the solution color to change from purple to yellow. This reaction leads to a decrease in absorbance at 517 nm, and the degree of decrease is positively correlated with the free radical scavenging ability of the antioxidant. Therefore, the DPPH method is widely used to evaluate the in vitro antioxidant activity of compounds or extracts.

[0099] In this embodiment, a 0.1 mM DPPH ethanol solution was prepared, and the antioxidant efficacy of the hair loss prevention and hair growth composition of the present invention was verified by utilizing the above-mentioned DPPH scavenging mechanism.

[0100] Using vitamin C as a positive control, 100 µL of the anti-hair loss and hair growth compositions prepared in Examples 1, 3, and 5 were respectively placed in a 96-well plate, 100 µL of DPPH ethanol solution was added, and after mixing, the mixture was reacted in the dark for 30 min. The absorbance value A was measured at a wavelength of 517 nm. s .

[0101] When an equal volume of anhydrous ethanol was used instead of the hair loss prevention and hair growth composition, the absorbance value A0 was measured under the same conditions; when an equal volume of anhydrous ethanol was used instead of the DPPH solution, the absorbance value A was measured under the same conditions. t .

[0102] DPPH removal rate is calculated using the formula: DPPH removal rate (%) = [1 - (A s -A t The calculation is performed using [) / A0]×100. The experiment was repeated three times, and the average value was taken. The test results are as follows: Figure 1 As shown.

[0103] Compared with the positive control system of vitamin C, the hair loss prevention and hair growth composition of the present invention exhibits significant antioxidant capacity, exceeding or reaching more than 80% of the efficacy of the positive control.

[0104] Example 10

[0105] Human dermal papilla cells (HDPs) are a type of specialized cell originating from the dermal connective tissue, located at the base of the hair follicle, and possessing unique spatial structure and physiological functions. HDPs activate the hair matrix cells surrounding them through paracrine signals, playing a central role in maintaining and regulating the hair follicle growth cycle.

[0106] After thawing frozen HDP cells, they were passaged in DEME high-glucose medium + 15% FBS complete medium to passages 3-5. Logarithmic growth phase cells were harvested, digested with 0.25% trypsin-0.02% EDTA, centrifuged at 1000 rpm for 5 min, and resuspended to a density of 2 × 10⁻⁶ cells / mL. 4 Cell suspension of cells / mL was seeded at 100 μL / well into 96-well plates and pre-cultured at 37°C, 5% CO2, and saturated humidity for 24 h. After the cells were fully adhered, the supernatant was discarded, and fresh culture medium containing gradient concentrations of the anti-hair loss and hair growth composition was added according to the experimental design. After culturing for another 24 h, cell activity was detected using the MTT assay to verify the proliferation-promoting effect of the anti-hair loss and hair growth composition of the present invention on HDP and to elucidate its mechanism of hair growth by activating key cell targets in hair follicles.

[0107] The core principle of the MTT assay relies on the activity of succinate dehydrogenase in the mitochondria of living cells. This enzyme catalyzes the reduction of yellow MTT molecules, producing water-insoluble blue-purple formazan crystals that deposit within the cells. Since dead cells lack this metabolic function and cannot produce crystals, the amount of formazan formed can indirectly reflect the number of living cells. The formazan crystals within the cells are dissolved in dimethyl sulfoxide (DMSO) to form a homogeneous purple solution. The absorbance of this solution is measured at 570 nm using a microplate reader, allowing for quantitative analysis of cell viability and proliferation.

[0108] Using the cells cultured above, add 20 μL of MTT solution to each well to achieve a final concentration of 0.5 mg / mL, and continue culturing for 3–4 h. Then, aspirate the culture medium from each well, add 150 μL of DMSO, and vortex at low speed for 10 min to dissolve the crystals. Measure the absorbance at 570 nm.

[0109] According to the formula: Cell viability (%) = (sample wells) OD - Withering Hole OD ) / (Blank control hole) OD - Withering Hole OD Cell viability was calculated by multiplying the result by 100%, and the results are as follows: Figure 2 As shown.

[0110] Figure 2 In the study, under gradient concentrations of 0.4%, 0.08%, and 0.0032% (v / v), the anti-hair loss and hair growth composition significantly enhanced the activity of HDP cells (110.30±1.74%~113.56±3.41%), showing a dose-dependent proliferative effect compared to the blank control group (100%). p <0.01).

[0111] Example 11

[0112] HDP cells cultured in Example 10 were treated with different concentrations of the anti-hair loss and hair growth composition for 24–48 hours. Cell supernatants were collected and the absorbance was measured at 450 nm according to the instructions of the TGF-β ELISA kit to verify the inhibitory effect of the anti-hair loss and hair growth composition of the present invention on TGF-β expression.

[0113] Simultaneously, a blank control group (BC) of untreated HDP cells and a positive control group (PC) of HDP cells treated with minoxidil were set up for comparison.

[0114] Based on the measurement results, the relative content of TGF-β and the inhibition rate of TGF-β were calculated according to the following formula:

[0115] Relative TGF-β content (%) = Average TGF-β content of sample group / Average TGF-β content of blank group × 100%;

[0116] TGF-β inhibition rate (%) = (relative content of sample group - relative content of blank group) / relative content of blank group × 100%.

[0117] like Figure 3 As shown, the anti-hair loss and hair growth composition significantly reduced the expression level of TGF-β at a low concentration, with effects comparable to the positive control group. Further analysis revealed that its inhibition rate of TGF-β increased with increasing concentration, exhibiting a typical dose-response relationship, consistent with the characteristics of active ingredients in pharmaceuticals and cosmetics.

[0118] Decreased TGF-β levels help slow down the apoptosis process of hair follicle cells, reverse the hair loss process, and thus effectively prevent hair loss and promote hair growth. These results confirm that the hair loss prevention and hair growth composition of this invention can effectively regulate TGF-β expression and has the potential to be used as a functional ingredient in hair loss prevention and hair growth drugs or cosmetics.

[0119] Example 12

[0120] Six-week-old male C57BL / 6 mice were acclimatized for one week under the following conditions: temperature 25±3℃, humidity 65%±5%, and 12h light-dark cycle. They were then randomly divided into a blank control group, a positive control group, and an experimental group, with six mice in each group.

[0121] Select a 2×3cm sample from the back of the mouse. 2 Hair removal is performed on the affected area using a razor and hair removal cream to completely remove hair. Medication is started on the day of hair removal (referred to as day 0).

[0122] The blank control group was treated with physiological saline, the positive control group was treated with 0.5% minoxidil, and the experimental group was treated with the anti-hair loss essence of Example 8 of this invention, with an application amount of 0.2 mL. The treatment was administered once daily for 18 consecutive days.

[0123] The hair growth in the bald areas of mice was observed and photographed daily. The hair regeneration status was recorded on days 1, 8, and 17 to evaluate the hair growth promoting effect of the hair loss prevention and hair growth composition of the present invention on mice.

[0124] Figure 4 The results showed that during the administration of the drug to mice, no redness, swelling, bleeding, or erosion occurred in any of the mice, indicating that the test samples did not cause any allergic reactions in the mice.

[0125] Hair follicle growth and development generally includes three main phases: the anagen (growth) phase, the catagen (transitional) phase, and the telogen (resting) phase. Under normal feeding conditions, C57BL / 6 mice typically enter the telogen phase at 6–8 weeks of age and do not spontaneously re-enter the anagen phase thereafter. In this experiment, by performing hair removal on the mice, the traumatic stimulation was used to induce the hair follicles to synchronously enter the anagen phase, subsequently experiencing the catagen and telogen phases in sequence.

[0126] Skin color changes in C57BL / 6 mice can reflect different hair growth cycles. When the skin is pink, the hair follicles are in the resting phase. When the hair follicles enter the growth phase from the resting phase, the melanocytes in the hair follicles secrete melanin, making the skin black. During the regression phase, melanin synthesis and secretion decrease, and the skin gradually turns gray until it enters the resting phase.

[0127] The hair growth of the mice in the three groups was observed. The hair growth of the mice in each group was consistent. Typical representative photos were selected as follows: Figure 4 As shown. On day 0, the skin of the shaved area of ​​the mouse is pink. On day 8, the skin on the back of the mouse begins to change from pink to black, that is, the hair follicles enter the growth phase. Hair begins to grow on day 10, and the hair is basically fully grown by day 17.

[0128] However, compared with the blank control group and the positive control group, it was clearly observed that the hair growth in the experimental group was vigorous and the hair was significantly longer than that in the other two groups. The hair was in a healthy growth state and there was no hair loss. The above results prove that the composition of the present invention has the effect of preventing hair loss and promoting hair growth.

[0129] Example 13

[0130] This embodiment uses the volunteer trial evaluation method to test the efficacy of the anti-hair loss and hair growth composition in accordance with the "Cosmetic Efficacy Evaluation Standard" and related human trial guidelines.

[0131] The anti-hair loss and hair growth shampoo of Example 7 and the anti-hair loss serum of Example 8 were used on 50-year-old male volunteers. The volunteers applied the anti-hair loss serum 2-3 times a week after shampooing, and continued use for 3 months. Subjective feedback from the volunteers was recorded regularly during the experiment, and comparison photos of hair growth were taken to objectively evaluate the effectiveness. Specific experimental results are as follows: Figure 5 As shown.

[0132] Based on consumer trial evaluation methods for the hair loss prevention and hair growth efficacy of cosmetics, combined with subjective feedback from volunteers, and Figure 5 Objective image analysis of the volunteers' hair showed that before use, the volunteers had large-scale hair loss on certain areas of their scalps; after one month of use, the volunteers reported a significant reduction in hair loss and some improvement in oil production; by the third month, large-scale hair loss had basically stopped, and the number of hairs had increased significantly.

[0133] Figure 5 The product also provides information on the condition of the scalp and hair follicles before and after use. Before use, the scalp showed an inflammatory reaction, which was indicated by a darker color in the image, with most hair follicles containing only one hair and excessive scalp oil secretion. After use, the scalp inflammation was significantly reduced, the number of hairs in each follicle increased to 2-3, scalp oil secretion decreased, and the overall scalp environment was improved.

[0134] It should be noted that this experiment was based on the synergistic effect of the anti-hair loss shampoo and anti-hair loss serum in terms of cleansing and conditioning, and the two were designed for combined use to better suit practical application scenarios. According to the product formulation design and previous research results, the anti-hair loss serum, as the carrier of the main active ingredients, has a certain improvement effect when used alone. When used together, it can synergistically regulate scalp sebum secretion and promote hair follicle health, thereby more comprehensively addressing hair loss problems and achieving better efficacy.

[0135] The above embodiments of the present invention do not describe all details exhaustively, nor do they limit the present invention to the embodiments described above. Various changes, modifications, substitutions, and variations made by those skilled in the art to these embodiments without departing from the principles and spirit of the present invention should be included within the scope of protection of the present invention.

Claims

1. A hair loss prevention and hair growth composition, comprising active ingredients and a solvent system: The active ingredients include watercress extract, peony root extract, ginkgo leaf extract, scutellaria root extract, sophora root extract, and Lactobacillus soyba fermentation product; The solvent system is an aqueous solution of butanediol and / or pentanediol; The above components are homogenized and compounded to form a composition that inhibits hair loss and promotes hair follicle activity.

2. The hair loss prevention and hair growth composition according to claim 1, characterized in that: The composition contains the following active ingredients by mass percentage: 1-20% watercress extract, 1-10% peony root extract, 0.1-0.3% ginkgo leaf extract, 0.1-0.3% scutellaria root extract, 0.5-1% sophora root extract, and 0.1-0.5% Lactobacillus soyba fermentation product.

3. The hair loss prevention and hair growth composition according to claim 1, characterized in that... The solvent system consists of butanediol and / or pentanediol, accounting for 20-30% of the composition by mass, and the balance being water.

4. The hair loss prevention and hair growth composition according to claim 1, characterized in that... Among them various The extract is obtained by extracting the active components with water or ethanol and then concentrating and freeze-drying them.

5. The hair loss prevention and hair growth composition according to claim 1, characterized in that... Soybeans Lactobacillus fermentation products are fermentation products with a solid content of ≥0.18wt% obtained by inoculating ground soybeans with lactobacillus strains and fermenting them.

6. The use of the anti-hair loss and hair growth composition of claim 1 as an active ingredient in the preparation of hair care products.

7. The application according to claim 6, characterized in that: The hair care products mentioned are anti-hair loss and hair growth shampoo, anti-hair loss conditioner / hair mask, anti-hair loss spray / serum, and anti-hair loss oil / essence.

8. The application according to claim 6, characterized in that: The hair loss prevention and hair growth composition is added to hair care products at a rate of 0.1 to 20 wt%.

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