Anti-hair-loss essence containing broadleaf holly leaf and Chinese ilex leaf extract and preparation method of anti-hair-loss essence

Abstract

The invention relates to an anti-alopecia essence containing a broadleaf holly leaf and Chinese ilex leaf extract and a preparation method thereof, the broadleaf holly leaf and Chinese ilex leaf extract is added into the anti-alopecia essence, and the anti-alopecia essence can show the effects of promoting hair growth and inhibiting alopecia after being used; the anti-hair loss essence further comprises a lactobacillus fermentation product and a penetration enhancer; as a conditioner, the lactobacillus fermentation product can improve the scalp microenvironment, and cooperates with the broadleaf holly leaf and Chinese ilex leaf extract to improve the alopecia condition; in addition, through the action of the penetration enhancer, the absorption degree of scalp to the broadleaf holly leaf and Chinese ilex leaf extract is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of anti-hair loss, and in particular relates to an anti-hair loss essence containing Ilex kudingcha leaves extract and a preparation method thereof. Background Art

[0002] Hair loss has become a global health challenge. With increasing stress and environmental pollution, the incidence of hair loss is rapidly increasing, and the disease is becoming more common among younger people. Consequently, demand for hair loss prevention and growth products is growing. Its pathogenesis is closely related to abnormal scalp lipid metabolism, oxidative stress damage, scalp microcirculatory disorders, and decreased hair follicle function.

[0003] There are two mainstream FDA-approved medications for the treatment of hair loss: topical minoxidil and oral finasteride. However, long-term use of minoxidil or finasteride is associated with a range of toxic side effects, including drug dependence, skin irritation, cardiovascular effects, and sexual dysfunction. Furthermore, given the complex etiology of hair loss, these two medications have a single mechanism of action and low transdermal efficiency, resulting in limited effectiveness in treating hair loss. Among non-drug treatments, plant extracts, due to their natural and safe properties, have become a research hotspot in the field of hair loss prevention and hair growth. Examples include ginseng, Polygonum multiflorum, and Platycladus orientalis leaves, which promote blood circulation in the scalp, enhance hair follicle nutrition, and inhibit fungal growth. These extracts can effectively improve the scalp environment and promote hair growth. However, the existing application of plant extracts still faces numerous challenges. For example, the extraction process is complex, the active ingredients are unclear or have poor stability, and some products have poor absorption, making them difficult to penetrate deeply into hair follicles. Furthermore, the efficacy or mechanism of action of some hair loss prevention products on the market lack scientific validation, or the products are expensive, failing to meet consumer demand. Therefore, there is an urgent need to develop safe and effective alternative therapies that utilize multiple synergistic mechanisms to address the current problem of hair loss.

[0004] Scalp essences, as important products for improving scalp conditions and promoting hair growth, have received widespread attention. However, existing scalp essence products have exposed many problems in actual use and cannot fully meet people's needs for a healthy scalp and strong, thick hair. From the perspective of scalp essence penetration, a large number of studies and actual usage feedback have shown that existing scalp essences generally have the problem of low penetration. The structure of the scalp is relatively complex, and its outermost layer, the stratum corneum, acts as a natural barrier, which has a certain obstructive effect on the absorption of substances. When using conventional scalp essences, the active ingredients in the essence often have difficulty effectively penetrating the stratum corneum and penetrating into the hair follicles and surrounding tissues where they work. From the perspective of conditioning the scalp environment and promoting hair growth, existing scalp essences also have obvious shortcomings. The scalp environment is a complex ecosystem, including multiple aspects such as microbial communities, oil secretion, moisture content, and acid-base balance.

[0005] A healthy scalp environment is fundamental to hair growth, yet many existing scalp serums fail to comprehensively and effectively regulate the scalp environment. Some scalp serums are designed to address only a specific scalp issue, such as simply controlling oil or moisturizing, while ignoring the holistic and complex nature of the scalp environment. For example, while some oil-control scalp serums can reduce oil secretion to a certain extent, they can also disrupt the scalp's water-oil balance, leading to excessive dryness, which in turn affects normal scalp metabolism and hair growth. Furthermore, existing scalp serums are ineffective in regulating the scalp's microbiome. The scalp is home to a variety of microorganisms, which normally exist in a state of equilibrium, working together to maintain scalp health. However, when the scalp microbiome is imbalanced, such as when Malassezia overgrows, it can cause itching and increased dandruff, which in turn affects hair follicle health and hair growth. Most current scalp serums lack ingredients or technologies that can effectively regulate the scalp microbiome balance, failing to fundamentally address scalp issues caused by microbial imbalance. Consequently, they struggle to create a favorable scalp environment conducive to hair growth, failing to achieve the goal of promoting hair growth. Summary of the Invention

[0006] In order to solve the above technical problems, the present invention provides an anti-hair loss essence containing Ilex kudingcha leaves extract and a preparation method thereof.

[0007] The technical solution adopted by the present invention is: an anti-hair loss essence containing Kudingcha Ilex leaf extract, comprising Kudingcha Ilex leaf extract and lactobacillus fermentation product; Kudingcha Ilex leaf is extracted by decocting in water, the filtrate obtained by decoction is concentrated and dried, and the obtained solid substance is Kudingcha Ilex leaf extract, and the Kudingcha Ilex leaf extract in the anti-hair loss essence accounts for 0.05%-10% by weight; the lactobacillus fermentation product is used as a conditioner, accounting for 0.1-10% by weight.

[0008] Preferably, the lactobacillus fermentation product is a lactobacillus fermentation lysate or a Lactobacillus plantarum fermentation product.

[0009] Preferably, the Lactobacillus fermentation product is replaced by Pichia pastoris fermentation lysate filtrate.

[0010] Preferably, the composition further comprises 2-8% penetration enhancer, 0.8-1.35% preservative, 2-15.5% moisturizer, 0.1-1% chelating agent, and the balance is solvent.

[0011] Preferably, the mass ratio of the Kudingcha Ilex leaf extract, the conditioner and the penetration enhancer is 1:2-7:4-10.

[0012] Preferably, the penetration enhancer is dimethyl isosorbide and / or ethoxydiglycol.

[0013] Preferably, the preservative is selected from one or more of methylparaben, caprylyl glycol, phenoxyethanol, ethylhexylglycerin, 1,2-hexanediol, and p-hydroxyacetophenone; The moisturizing agent is selected from one or more of glycerin, D-panthenol, propylene glycol, butylene glycol, and betaine; The solvent is water or a mixture of water and ethanol.

[0014] A method for preparing an anti-hair loss essence containing Ilex kudingcha leaf extract comprises adding a solvent, Ilex kudingcha leaf extract, a preservative, a moisturizer, and a chelating agent into an emulsifying tank, starting stirring at a speed of 20-40 rpm, heating to 80° C. and maintaining the temperature for 10-15 minutes; Stir and cool down to 40-50°C, add conditioner, stir evenly until the temperature is below 40°C, remove from the pot, and age for 48 hours to obtain the anti-hair loss essence containing Kudingcha Ilex leaf extract.

[0015] Preferably, the pH value of the anti-hair loss essence is adjusted to 5.0-7.0; Preferably, the pH value of the anti-hair loss serum is adjusted by lactic acid, citric acid or arginine.

[0016] Preferably, the Kudingcha Ilex leaf extract includes Kudingsaponin active substances.

[0017] The advantages and positive effects of the present invention are as follows: the scalp essence containing the Kudingcha Ilex leaf extract can promote hair growth and inhibit hair loss; the addition of lactobacillus fermentation products as a conditioning agent can improve the scalp microenvironment and synergistically improve hair loss with the Kudingcha Ilex leaf extract; and the absorption of the Kudingcha Ilex leaf extract and the lactobacillus fermentation products by the scalp is improved through the action of a penetration enhancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Comparison of the proliferation activity of active substances from Ilex kudingcha L. leaves on hair follicle stem cells; Figure 2 Mass spectrometry analysis of total saponins in Ilex kudingcha leaf extract; Figure 3 Determination of total saponin content in extracts of Ilex kudingchaenensis, Ilex macrophylla and Ilex oleifera; Figure 4 Comparison of the proliferation activity of total saponins from Ilex kudingchaenensis, Ilex macrophylla and Ilex oleraceus on hair follicle stem cells; Figure 5 Mass spectra of holly saponin components in Ilex kudingchaenensis, Ilex macrophylla and Ilex oleraceus; Figure 6 Figure 3: The effect of Ilex kudingcha L. extract on preventing hair loss in mice with androgenic alopecia; Figure 7Effects of different conditioning agents on the regulation of scalp microenvironment by anti-hair loss essence containing Ilex kudingcha leaves extract; Figure 8 Effects of Lactobacillus plantarum fermentation products on the regulation of scalp microenvironment by anti-hair loss essence containing Ilex kudingcha leaves extract; Figure 9 The effect of the anti-hair loss essence containing Ilex kudingcha leaf extract on androgenic alopecia mice; Figure 10 Hair mirror analysis before and after using the anti-hair loss essence containing Ilex kudingcha leaf extract. DETAILED DESCRIPTION

[0019] The embodiments of the present invention are described below with reference to the accompanying drawings.

[0020] The present invention relates to an anti-hair loss essence containing a Kudingcha Ilex leaf extract and a preparation method thereof. A plant extract is added to a hair care product essence, wherein the plant extract is a Kudingcha Ilex leaf extract or a plant composition containing Kudingcha Ilex leaf as a main ingredient. The anti-hair loss essence can promote hair growth and inhibit hair loss after use.

[0021] Kudingcha Holly is a tree plant of the genus Ilex of the Aquifoliaceae family, mainly produced in Hainan and Guangxi. Its dried leaves are mainly used as medicine and food. It is cool in nature, bitter in taste, and slightly sweet in flavor. It has the effects of dispelling wind and heat, clearing the head, relieving restlessness and quenching thirst. Kudingcha has a long history in my country. The representative varieties are large-leaf holly ( Ilex latifolia Thunb.), Ilex kudingcha ( Ilex kudingcha CJ Tseng) and holly ( Ilex cornuta Lindl. et Paxt.). The main component of Ilex kudingcha is saponin, and it also contains flavonoids, polyphenols, polysaccharides, and volatile oils. Extracts obtained by extracting the active ingredients from Ilex kudingcha leaves can be used to prepare topical preparations for preventing hair loss.

[0022] The Kudingcha Ilex leaf extract is obtained by extracting the Kudingcha Ilex leaf by water decoction. The solution after water decoction extraction can be further concentrated and dried, and the obtained Kudingcha Ilex leaf extract can be used as a raw material for the preparation of other products. In certain embodiments of the present invention, the Kudingcha Ilex leaf extract is an effective component including total polysaccharides, total phenolic acids, total saponins, and total flavonoids. The total polysaccharides, total phenolic acids, total saponins, and total flavonoids in the Kudingcha Ilex leaf water extract can be separated by different concentrations of ethanol. The separated total polysaccharides, total phenolic acids, total saponins, and total flavonoids are then mixed and compounded to obtain a new type of Kudingcha Ilex leaf extract. In other embodiments of the present invention, the Kudingcha Ilex leaf extract specifically refers to the extracted saponin active substances.

[0023] The Kudingcha Ilex leaves are boiled with water, and distilled water is added to the Kudingcha Ilex leaves at a w / v ratio of 1:10-15 to decoct the mixture to extract the active ingredients. The extraction time is 0.5-3 hours, and the filtered solution is concentrated and dried to obtain the Kudingcha Ilex leaf extract. Alternatively, the Kudingcha Ilex leaves are boiled with water to extract the active ingredients, the filtrate is concentrated, and 70% ethanol is added to the precipitate to obtain the total polysaccharides. The supernatant is extracted and loaded onto an adsorption resin column. The components eluted with 30% ethanol are the total phenolic acids, the components eluted with 50% ethanol are the total saponins, and the components eluted with 80% ethanol are the total flavonoids. The total polysaccharides, total phenolic acids, total saponins, and total flavonoids are mixed and dried to obtain the Kudingcha Ilex leaf extract. Alternatively, only the total saponin component prepared in the aforementioned method is used as the Kudingcha Ilex leaf extract; 70% ethanol is added to the aqueous extract of Kudingcha Ilex leaf to remove total polysaccharides, the separated liquid portion is loaded onto an adsorption resin column, eluted with 30% ethanol to remove total phenolic acids, and then the 50% ethanol elution component is collected, concentrated and dried to obtain the total saponins of Kudingcha Ilex leaf.

[0024] Experimental validation has shown that Ilex kudingcha significantly suppresses triglyceride, total cholesterol, and 5α-reductase levels in a mouse model of androgenic alopecia. It also inhibits elevated levels of ROS or MDA, or abnormal expression of inflammatory factors such as TNF-α, IL-1β, IL-6, and INF-γ, in mouse hair follicle stem cells induced by ultraviolet irradiation or hydrogen peroxide. Ilex kudingcha leaf extract can be used to improve hair loss caused by various mechanisms, such as inhibiting abnormally elevated oil secretion in the scalp sebaceous glands or excessive androgen levels, eliminating hair loss caused by elevated levels of reactive oxygen species (ROS) in the scalp induced by physical damage or chemical stimulation, and inhibiting hair loss caused by abnormal expression of inflammatory factors in the scalp induced by physical, chemical, or biological factors.

[0025] Adding Kudingcha Ilex leaf extract to a scalp serum in a hair care product can enhance the Kudingcha Ilex leaf extract's effects in promoting hair growth and inhibiting hair loss when used in the scalp serum. Further, the synergistic effect of additives such as conditioners and penetration enhancers can further enhance its beneficial effects in controlling hair loss. By weight, the Kudingcha Ilex leaf extract accounts for 0.05%-10% of the scalp serum. In certain embodiments of the present invention, the hair care product containing Kudingcha Ilex leaf extract also includes a penetration enhancer, a conditioner, a preservative, a moisturizer, a chelating agent, and a solvent. By weight, the composition includes 2-8% of a penetration enhancer, 0.1-10% of a conditioner, 0.8-1.35% of a preservative, 2-15.5% of a moisturizer, 0.1-1% of a chelating agent, and the remainder is solvent.

[0026] The penetration enhancer is a combination of one or both of isosorbide dimethyl ether and ethoxydiglycol; the conditioning agent is lactobacillus ferment or Pichia pastoris fermentation lysate filtrate, and the lactobacillus ferment can be specifically lactobacillus ferment lysate or plantarum ferment; the preservative is selected from one or more of methylparaben, octoxyglycerin, phenoxyethanol, ethylhexylglycerin, 1,2-hexanediol, and p-hydroxyacetophenone; the humectant is selected from one or more of glycerin, D-panthenol, propylene glycol, butylene glycol, and betaine; and the chelating agent is EDTA-2Na.

[0027] To prepare the anti-falling essence, the solvent, the Ilex kudingcha C.J. Tseng and Cyclobalanopsis glauca extract, the preservative, the humectant, and the chelating agent are added to an emulsifying tank, and stirring is started at a speed of 20-40 rpm; the temperature is raised to 80°C and kept for 10-15 min; the stirring is started to reduce the temperature to 40-50°C, the conditioning agent is added, the pH value of the anti-falling essence is adjusted to 5.0-7.0 by lactic acid, citric acid, or arginine, and the stirring is uniform until the temperature is below 40°C; the tank is emptied, and the anti-falling essence containing the Ilex kudingcha C.J. Tseng and Cyclobalanopsis glauca extract is obtained after 48 hours of storage.

[0028] The anti-inflammatory and oil-controlling core effects of Ilex kudingcha C.J. Tseng depend on the active ingredients reaching the deep target points of the scalp (such as hair follicles, sebaceous glands, and inflammatory cells), but the barrier effect of the scalp stratum corneum and hair follicle openings hinders the penetration, resulting in most of the ingredients only staying on the surface and having limited effects. The penetration enhancer can help the active ingredients of Ilex kudingcha C.J. Tseng penetrate the barrier by opening the stratum corneum channels, directly reaching the dermis layer and the surrounding hair follicles, so that the anti-inflammatory effect is upgraded from “surface soothing” to “deeply inhibiting inflammatory factors (such as IL-6 and TNF-α)”, and the oil-controlling effect is changed from “temporary adsorption of oil” to “inhibition of excessive secretion of sebaceous gland cells”, greatly improving the improvement efficiency of scalp problems (such as dandruff, itching, and greasiness). The penetration enhancer promotes the penetration and absorption of active substances by changing the structure of the stratum corneum, increasing the solubility of the active substances, changing the protein structure to enhance the sebaceous gland pathway, or increasing the water content; and a scientific and effective penetration mechanism is established by matching the types and proportions of the conditioning agent and the penetration enhancer.

[0029] Furthermore, if the scalp barrier is damaged, the disordered stratum corneum can lead to unstable penetration of the penetration enhancer. Furthermore, the scalp beneath this damaged barrier is susceptible to external irritants, and the anti-inflammatory properties of Kudingcha tea may be overwhelmed by superficial irritants, preventing them from deeply exerting their effects. Conditioners stabilize the scalp's physiological environment by repairing the barrier and maintaining the water-oil balance. This not only allows for more controlled penetration of the penetration enhancer, but also provides a direct path for Kudingcha tea's active ingredients to reach their targets. The penetration enhancer breaks down the penetration barrier, amplifying Kudingcha tea's anti-inflammatory and oil-control properties and the conditioning agent's moisturizing and repairing properties. Kudingcha tea protects other ingredients through its antioxidant and natural anti-inflammatory properties, enhancing their targeted effects. The conditioning agent repairs the barrier, stabilizes the penetration environment, and reduces the risk of irritation. By optimally combining Kudingcha ilex leaf extract, the penetration enhancer, and the conditioning agent, and through compatibility screening, the synergistic effects of these three can be maximized, achieving a comprehensive scalp health management approach characterized by "deep repair + precise oil control + barrier stabilization."

[0030] The present invention is described below with reference to the accompanying drawings. Experimental methods without specific operating steps are carried out in accordance with the corresponding product specifications. Unless otherwise specified, the instruments, reagents, and consumables used in the examples can be purchased from commercial companies.

[0031] Example 1: Preparation and characterization of Ilex kudingcha leaf extract

[0032] 1.1 Preparation of Ilex kudingcha L. Leaf Extract

[0033] The dried Ilex kudingcha leaves are taken, crushed and then boiled in water to extract the active ingredients of Ilex kudingcha leaves.

[0034] 1000 g of dried Ilex kudingcha leaves were weighed and crushed, and then heated and boiled with distilled water at a w / v ratio of 1:15 for 2 h. The mixture was filtered while hot using a 100-mesh stainless steel sieve, and the filtrate was concentrated using a flash evaporator (JMF-320, Xi'an). The concentrated product was dried to obtain the solid substance, which was the Ilex kudingcha leaf extract.

[0035] 1.2 Analysis of components of Ilex kudingcha leaf extract

[0036] In order to further analyze the key active ingredients in the leaves of Ilex kudingcha, different categories of active ingredients in the extract of Ilex kudingcha leaves were separated and purified, and total polysaccharides, total phenolic acids, total saponins and total flavonoids were collected respectively, and the functions of various active ingredients were analyzed and verified.

[0037] 1000 g of dried Ilex kudingcha leaves were weighed and crushed. Extraction was then heated and boiled with distilled water at a w / v ratio of 1:15 for 2 h. The extract was filtered hot through a 100-mesh stainless steel sieve, and the filtrate was concentrated to a suspension with a density of 1.1 (approximately 2.5 L) using a flash evaporator (JMF-320, Xi'an). Ethanol was added to the Ilex kudingcha leaf extract suspension to adjust the ethanol content to 70%, mixed thoroughly, and allowed to stand overnight. The precipitate was washed three times with anhydrous ethanol, resulting in a total of 35.4 g of total polysaccharides, the main component. The supernatant was further concentrated to a solution with a solids content of approximately 1%. The solution was loaded onto an AB-8 macroporous adsorption resin column (diameter: 5.5 cm; column height: 35 cm, column volume: approximately 850 mL) at a flow rate of 2 mL / min. The column was equilibrated with 5% ethanol to remove water-soluble impurities. The elution was then performed using a gradient elution method with varying ethanol concentrations at a rate of 1 mL / min. Among them, the components obtained from the 30% ethanol solution elution part were mainly total phenolic acids, totaling 18.7 g; the components obtained from the 50% ethanol solution elution part were mainly total saponins, totaling 21.8 g; and the components obtained from the 80% ethanol solution elution part were mainly total flavonoids, totaling 28.4 g.

[0038] 1.3 Effects of active ingredients from Ilex kudingcha leaves on the proliferation of hair follicle stem cells

[0039] The isolated total polysaccharides, total phenolic acids, total saponins, and total flavonoids were tested for the proliferation activity of rat hair follicle stem cells (HFSCs). Rat hair follicle stem cells (HFSCs) were purchased from Wuhan Punosai Life Science Technology Co., Ltd. HFSCs were placed in a dedicated culture medium and cultured continuously in a 37°C, 5% CO2 incubator. The medium was changed every other day. When the cells reached 70% to 80% confluency, HFSCs in the logarithmic growth phase were passaged and cultured at a rate of (1×10 4 100 μL of the culture medium (per well) was inoculated into a 96-well plate, and a control group, Ilex kudingcha group, total polysaccharide group, total phenolic acid group, total saponin group, and total flavonoid group were set up. The control group was a basal culture medium, and the other groups were basal culture medium solutions containing 0.1% of the corresponding components. HFSCs were inoculated into the culture medium of the control group and each experimental group, and continued to be cultured in the incubator for 24 h. 10 μL of CCK-8 reagent was added to each group, and the cells were returned to the incubator for continued culture for 1 h. The absorbance value was detected at 450 nm. Three replicate wells were set up for each group to ensure the accuracy of the experiment. The growth rate of cell proliferation in each experimental group was compared with that of the control group, and the cell proliferation enhancement rate was calculated to measure the promoting effect of different experimental groups on cell proliferation. Cell proliferation enhancement rate % = (absorbance value of experimental group - absorbance value of control group) / absorbance value of control group × 100%.

[0040] Depend on Figure 1 The results showed that the Ilex kudingcha leaf extract significantly promoted the proliferation of hair follicle stem cells (HFSCs), increasing the average cell proliferation rate by 37.1% compared to the control group. Its main components (total polysaccharides, total phenolic acids, total saponins, and total flavonoids) all exhibited some proliferative activity, with the total saponins group showing the most significant effect (increasing the proliferation rate by 58.4%). This suggests that Ilex kudingcha has the ability to enhance the activity of hair follicle stem cells, suggesting its potential application in preventing hair loss, with total saponins representing the core active components.

[0041] 1.4 Analysis and performance comparison of total saponins in Ilex kudingcha leaf extract

[0042] The key components of the total saponins, the key active ingredients in the leaves of Ilex kudingcha, were further identified and analyzed, and the components were separated and identified by mass spectrometry analysis using UPLC-Q-TOF / MS.

[0043] Chromatographic column: CORTECS T3 (2.1 mm × 150 mm, 1.6 μm); chromatographic gradient elution conditions: mobile phase A: acetonitrile, mobile phase B: 0.1% formic acid in water; flow rate: 0.2 mL / min; gradient elution (0-5 min: 5% A, 95% B; 5-35 min: 25%, 75% B; 35-40 min: 40% A, 60% B; 40-45 min: 75% A, 25% B; 45-50 min: 95% A, 5% B). The results are shown in Figure 5. Figure 2 The figure shows a section of the key saponin components, and the most representative saponin components were identified through MS / MS fragment analysis, as shown in Table 1. The main components are kudingcha holly saponins, namely kudinoside G, kudinoside C, kudinoside A, and kudinoside D. Fractions were collected according to their peak elution time, and the corresponding four saponin components were enriched, concentrated, and dried. The fraction from 36.5 min to 40.41 min was also collected, concentrated, and dried. The hair follicle stem cell proliferation-promoting activity of the four kudingcha holly saponins was assayed according to the method described in 1.3. The results are shown in Table 1. All four saponin components exhibited hair follicle stem cell proliferation-promoting activity at a concentration of 10 μg / mL, but the activity was lower than that of the fraction from 36.5 min to 40.41 min (total kudingcha holly saponins) at the same concentration. Comparison revealed that a mixture of different kudingcha holly saponin components (total kudingcha holly saponins) exhibited a more potent effect.

[0044] Table 1 Analysis of total saponins and their activity in Ilex kudingcha leaves extract

[0045]

[0046] Example 2: Determination of total saponin content and performance analysis in Ilex kudingcha, Ilex glabra and Ilex oleifera

[0047] 2.1 Comparison of total saponin content in Ilex kudingchaenensis, Ilex macrophylla, and Ilex oleraceus

[0048] Representative Kudingcha varieties, including Ilex kudingcha, Ilex oleracea, and Ilex oleracea, were compared. These Ilex kudingcha, Ilex oleracea, and Ilex oleracea leaves were purchased from medicinal herb markets in Guangxi Zhuang Autonomous Region, Zhejiang Province, and Guizhou Province, respectively, and their total saponin content was determined.

[0049] The total saponin content of three batches of Ilex kudingchaenensis, Ilex glacialis, and Ilex oleifera was determined using UV-visible spectrophotometry, using ginsenoside Re (purchased from Shanghai Yuanye Biotechnology Co., Ltd., Catalog No. A10036) as a reference and vanillin-glacial acetic acid as a colorimetric method. Specifically, 100 μL of the test or reference solution was accurately pipetted into a 10 mL stoppered test tube. The solvent was evaporated in a 60°C water bath. 0.2 mL of a 10% vanillin-glacial acetic acid solution and 0.8 mL of perchloric acid were added. The tube was incubated in a 60°C water bath for 15 minutes, then placed in an ice-water bath for 5 minutes. 5 mL of glacial acetic acid was added, mixed, and the absorbance was measured at 550 nm.

[0050] Preparation of reference solution: Accurately weigh 10 mg of ginsenoside Re reference solution and place it in a 10 mL volumetric flask. Add methanol to the volume and shake well to obtain a 1 mg / mL ginsenoside Re reference solution.

[0051] Accurately pipette 0, 10, 20, 40, 60, 80, and 100 μL of ginsenoside Re reference solution (1 mg / mL). Measure the absorbance A value according to the above method. Plot a standard curve with A mass concentration as the abscissa (X) and A value as the ordinate (Y). The resulting linear regression equation is y = 0.0204x + 0.0017, R 2 =0.9995.

[0052] Preparation of test solution: Weigh 100 g each of dried Ilex kudingcha leaves, Ilex macrophylla leaves, and Ilex rutaecarpa leaves, add 10 times the volume of water, soak for 30 minutes, heat to boiling, and extract for 1 hour. Filter, concentrate the filtrate to a relative density of 1.1-1.2, dry under reduced pressure, and pulverize to obtain the extract. Accurately weigh 10 mg of each extract powder and place it in a 10 mL volumetric flask. Add methanol to the volume and shake thoroughly to obtain the test solution.

[0053] The total saponin content in the test samples was determined according to the above method, and the results are shown in Table 2. Among them, the total saponin content of Ilex kudingcha was the highest, with an average content of 7.43%; the total saponin content of Ilex kudingcha was 5.18%; the total saponin content of Ilex kudingcha was 4.96%. Figure 3 As shown in the figure, it can be seen that Ilex kudingcha has a higher content of active ingredient total saponins than the other two types.

[0054] Table 2 Total saponin contents of three extracts from different batches of Ilex kudingcha, Ilex macrophylla and Ilex oleraceus

[0055]

[0056] 2.2 Comparison of the proliferation activity of total saponins from Ilex kudingchaenensis, Ilex grandiflora, and Ilex oleraceus on hair follicle stem cells

[0057] According to the preparation method of total saponins in Example 1, three total saponins of Ilex kudingchai, Ilex grandiflora and Ilex scoparia were separated respectively, and the effects of the three total saponins on the proliferation activity of hair follicle stem cells were determined at a concentration of 0.1%.

[0058] HFSCs cells were passaged when the confluence reached 70% to 80%. HFSCs in the logarithmic growth phase were selected and cultured according to the (1×10 4 100 μL of the solution (100 μL per well) was inoculated into a 96-well plate. Three groups were set up: a control group (Ilex kudingcha), a group with Ilex kudingcha (Ilex kudingcha), a group with Ilex larvae (Ilex larvae), and a group with Holly. The control group used basal medium, while the other groups all used basal medium supplemented with 0.1% of the corresponding total saponins. The Ilex kudingcha group consisted of one-third Ilex kudingcha, one-third Ilex larvae, and one-third Holly. The plates were incubated in an incubator for 24 hours. Then, 10 μL of CCK-8 reagent was added and the plates were returned to the incubator for another hour. The absorbance was measured at 450 nm. Three replicate wells were set up for each group to ensure experimental accuracy.

[0059] The cell proliferation enhancement rate was calculated according to the method in Example 1. The results were as follows: Figure 4 As shown in the results, the total saponins from Kudingcha (Ilex kudingcha), Ilex kudingcha (Ilex kudingcha), Ilex kudingcha (Ilex kudingcha), and Ilex kudingcha (Ilex kudingcha) groups all significantly promoted the proliferation of HFSCs. Compared with the control group, the average cell proliferation rate increased by 41.1% for Kudingcha total saponins, 36.8% for Ilex kudingcha, 27.9% for Ilex kudingcha, and 57.9% for Ilex kudingcha (Ilex kudingcha). This indicates that at the same level, the activity of total saponins from Ilex kudingcha (Ilex kudingcha) was significantly higher than that from Ilex kudingcha, Ilex kudingcha, or the combination of the three (Kudingcha), indicating that Kudingcha (Ilex kudingcha) may have the greatest potential for anti-hair loss among all Kudingcha teas.

[0060] 2.3 Analysis of saponin components in Ilex kudingchaenensis, Ilex macrophylla, or Ilex oleraceus

[0061] Furthermore, the key components of the total saponins, which are the key active ingredients of Ilex kudingchai, Ilex grandiflora or Ilex oleifera, were identified and compared, and the components were separated and identified by mass spectrometry analysis using UPLC-Q-TOF / MS.

[0062] According to the mass spectrometry analysis conditions shown in Example 1, three samples of 10 mg / mL were tested in parallel. The test results are as follows: Figure 5 As shown in the figure, the three ingredients share similarities in their component categories, while also exhibiting significant differences in their content. Comparing the relative peak areas, we calculated that the content of kudinoside G, kudinoside C, kudinoside A, and kudinoside D in Ilex kudingchaenensis was 1.57 times that of Ilex leucophylla and 2.54 times that of Ilex oleraceus. This indicates that the content of active holly saponins in Ilex kudingchaenensis is significantly higher than that in Ilex leucophylla or Ilex oleraceus.

[0063] The fractions from 36.5 min to 40.41 min were collected and concentrated, and the hair follicle stem cell proliferation-promoting activity of the four Ilex kudingchai saponins was assayed according to the method described in Example 1. The results are shown in Table 3. It can be seen that the saponins from Ilex kudingchai, Ilex euphorbiae, or Ilex oleracea all exhibited hair follicle stem cell proliferation-promoting activity at a concentration of 10 μg / mL, but there were significant differences in activity. The Ilex kudingchai saponins exhibited the highest activity, significantly superior to those from Ilex euphorbiaee or Ilex oleracea. This indicates that different total saponin extracts exhibit varying abilities to promote hair follicle stem cell proliferation due to differences in the ratios of their saponin components.

[0064] Table 3 Proliferative activity of holly saponins from Ilex kudingchaenensis, Ilex macrophylla, or Ilex oleraceus

[0065]

[0066] Example 3: Preparation of scalp essence containing Ilex kudingcha leaves extract

[0067] Add the solvent, Ilex kudingcha leaf extract, preservative, moisturizer, and chelating agent into an emulsifying tank, start stirring at a speed of 20-40 rpm; raise the temperature to 80°C and keep it warm for 10-15 minutes; start stirring and cool down to 40-50°C, add a conditioner, adjust the pH value of the scalp essence to 5.0-7.0 with lactic acid, stir evenly until the temperature is below 40°C, remove from the pot, and age for 48 hours to obtain a scalp essence containing Ilex kudingcha leaf extract.

[0068] Example 4: Effect of Ilex kudingcha L. Extract Content on Scalp Essence

[0069] To investigate the anti-hair loss efficacy of Ilex kudingcha leaf extract, the therapeutic effects of scalp extracts supplemented with different ratios of Ilex kudingcha leaf extract on an androgenic alopecia model mouse model were compared. Scalp extracts containing 0.05, 1, 3.35, 5, and 10 weight percent Ilex kudingcha leaf extract (Experiments 1-2, 1-3, 1-4, 1-5, and 1-6) were prepared according to the method of Example 3. Additionally, a scalp extract without Ilex kudingcha leaf extract was prepared as Experimental Example 1-1, and also contained 0.2% methylparaben, 0.35% phenoxyethanol, 0.8% ethylhexylglycerin, 0.3% D-panthenol, 5% butylene glycol, 0.2% betaine, and 0.5% EDTA-2Na, with the balance being water.

[0070] 40 male C57BL / 6J mice were randomly divided into 8 groups: blank group, model group, experimental example 1-1, experimental example 1-2, experimental example 1-3, experimental example 1-4, experimental example 1-5 and experimental example 1-6. Except for the blank group, the other groups were evenly applied with 200 μL of testosterone (5 mg / mL) every day for modeling, and 200 μL of the test substance was applied after complete absorption. The model group was treated with a blank matrix of essence without active substances for 14 consecutive days. The tape stripping test is used to evaluate the firmness of the hair. After the experimental mice were anesthetized, the hair growth in the hair loss area on the back of the mice was photographed, and then a 1cm 2 A hair loss test was performed on the back test area using transparent tape. The number of hairs removed was counted to reflect the anti-hair loss effect of each group. Any hair that grew in the shaved area was shaved with a shaver, collected with weighing paper, and weighed using a 1 / 100,000 electronic balance.

[0071] Effects of Ilex kudingcha on the amount and weight of hair loss in mice with androgenic alopecia Figure 6 The results show that Experimental Example 1-1 had no significant effect on preventing hair loss, and the mice's hair remained consistent with the model group. The Kudingcha Ilex leaf extracts from Experimental Examples 1-2 to 1-6 significantly inhibited hair loss in male mice, reducing the number of hair loss and increasing the weight of hair in the affected areas. The effect of the treatment group in Experimental Example 1-3 (containing 1% Kudingcha Ilex leaf extract) was the best, along with the treatment groups in Experimental Example 1-4 (containing 3.35% Kudingcha Ilex leaf extract), Experimental Example 1-5 (containing 5% Kudingcha Ilex leaf extract), and Experimental Example 1-6 (containing 10% Kudingcha Ilex leaf extract), all demonstrating significant anti-hair loss effects.

[0072] Example 5: Evaluation of the Effect of Penetration Enhancers on the In Vitro Release and Transdermal Properties of Kudingcha Scalp Essence

[0073] To maximize the effectiveness of Ilex kudingcha leaf extract, it was formulated with a penetration enhancer, such as dimethyl isosorbide and / or ethoxydiglycol. To investigate the effects of different types and amounts of penetration enhancers on the in vitro release and transdermal properties of the active ingredients in Ilex kudingcha scalp extract, Ilex kudingcha leaf extract scalp extracts containing different types and amounts of penetration enhancers were prepared. The composition and content (weight percentage) of the penetration enhancers are shown in Table 4.

[0074] Table 4

[0075]

[0076] The scalp essence also includes 1% Ilex kudingcha leaf extract, 0.2% methylparaben, 0.35% phenoxyethanol, 0.8% ethylhexylglycerin, 0.3% D-panthenol, 5% butylene glycol, 0.2% betaine and 0.5% EDTA-2Na, with the balance being water.

[0077] The permeation performance of each group of scalp essences was compared. An in vitro permeation test was designed based on the guidelines of the Pharmaceuticals and Medical Devices Agency (PMDA) of Japan. A porcine skin / Franz diffusion cell permeability test system was constructed. The permeation test was conducted to investigate the transdermal behavior of Kudingcha scalp essences containing different types and amounts of permeation enhancers in piglet skin.

[0078] First, seven pieces of Bama Xiang pig back skin were prepared. Using physiological saline as the receiving solution, the skins were mounted on a Franz diffusion cell. The receiving solution was added to the lower chamber, and stirring and temperature control were initiated. The cells were equilibrated at 350 rpm and 32±1°C for 30 min. Samples from Experiments 2-1 to 2-7 were then dripped onto the skins, with a 1 mL sample volume. The upper chamber was sealed with parafilm for testing. After 12 h, all samples were aspirated with the receiving solution and the skins were removed. The receiving solution was filtered through a 0.22 μm microporous membrane for subsequent testing. The percutaneous area of ​​the skin was excised with surgical scissors and washed three times with physiological saline. Filter paper was used to wipe the surface of the skin to remove any residual sample. After washing, three times the volume of methanol was added, the cells were minced, sonicated, and centrifuged. The supernatant was collected for subsequent testing.

[0079] UV-visible spectrophotometry was used to determine the content of total saponins from Ilex kudingchaenensis in pig skin and the receiving solution using ginsenoside Re (purchased from Shanghai Yuanye Biotechnology Co., Ltd., Catalog No. A10036) as a reference. The vanillin-glacial acetic acid method was used for color development, and its in vitro transdermal behavior was characterized. Specifically, 100 μL of the test or reference solution was precisely pipetted into a 10 mL stoppered test tube. The solvent was evaporated in a 60°C water bath. 0.2 mL of a 10% vanillin-glacial acetic acid solution and 0.8 mL of perchloric acid were added. The tube was incubated in a 60°C water bath for 15 minutes, then placed in an ice-water bath for 5 minutes. 5 mL of glacial acetic acid was added, mixed, and the absorbance was measured at 550 nm.

[0080] The effects of different penetration enhancers on the in vitro release and transdermal properties of Kudingcha scalp essence are shown in Table 5. It can be seen that penetration enhancers can have different promoting effects on the scalp of Kudingcha Ilex oleifera, among which Experimental Example 2-3 has the best transdermal absorption effect, with a skin permeation rate of 44.17%.

[0081] Table 5 Effects of different types and amounts of penetration enhancers on the skin permeability of Kudingcha Ilex oleifera essence

[0082]

[0083] Example 6: Experimental evaluation of the effect of conditioners on the scalp microenvironment

[0084] The scalp microenvironment, also known as the scalp microbiome, is composed of host cells and their secretions on the scalp surface, scalp microbes, and the microenvironment they inhabit. When the scalp microenvironment is imbalanced, scalp microbes and their secretions can induce epidermal hyperkeratosis and excessive sebum secretion, leading to cell apoptosis, increased chemokine recruitment, and increased expression of invasive enzymes. These factors ultimately infiltrate inflammatory cells surrounding hair follicles, altering the scalp's normal pH and disrupting the scalp barrier, triggering symptoms such as scalp inflammation, oily scalp, itching, and redness, ultimately leading to hair loss.

[0085] A conditioning agent may be further added to the scalp serum containing Ilex kudingcha leaf extract.

[0086] Conditioners were Lactobacillus fermentation lysate (Shanghai Baihaobo Biotechnology Co., Ltd., batch number: LDHG-010723), Pichia pastoris fermentation lysate filtrate (PFLF2) (Guangzhou Xiandi Biotechnology Co., Ltd., batch number: 20240304-1), or Lactobacillus plantarum fermentation product. The Lactobacillus plantarum fermentation product was prepared by inoculating a small amount of Lactobacillus plantarum from a laboratory collection onto a solid plate. Using the streak inoculation method, the strain was spread onto the solid plate and incubated at 37°C in a constant temperature incubator for 24–36 hours to allow the strain to grow and form single colonies on the solid plate. Well-grown colonies were selected and added to 10 mL of liquid MRS medium. The culture was incubated for 12–14 hours, and then filtered through a 0.22 μm pore size filter to remove residual bacteria and impurities to obtain the Lactobacillus plantarum fermentation product.

[0087] Lactobacillus plantarum is selected from Lactobacillus plantarum 18 or 24. Lactobacillus plantarum 18 is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with a deposit number of CGMCC No. 31277, a deposit date of July 12, 2024, and a classification name of Lactobacillus plantarum. Lactiplantibacillus plantarum , deposit address: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing; Lactobacillus plantarum 24 is deposited in the General Microbiology Center of China Culture Collection Administration, deposit number: CGMCC No. 31278, deposit date: July 12, 2024, classification name: Lactobacillus plantarum Lactiplantibacillus plantarum , Collection address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0088] In order to explore the effects of different conditioners on the scalp microenvironment, the above-mentioned fermentation products containing 0.1% of different types of conditioners were prepared, including Experimental Example 3-1-1 (Lactobacillus fermentation lysate), Experimental Example 3-1-2 (Pichia pastoris fermentation lysate filtrate-PFLF2) and Experimental Example 3-1-3 (Lactobacillus plantarum fermentation product), which also included 1% Ilex kudingcha leaf extract, 5% dimethyl isosorbide, 0.2% methylparaben, 0.35% phenoxyethanol, 0.8% ethylhexylglycerin, 0.3% D-panthenol, 5% butylene glycol, 0.2% betaine and 0.5% EDTA-2Na, and the balance was water.

[0089] Appropriate volunteers were recruited for a hair loss prevention test using Kudingcha Wintergreen Extract. Eighteen volunteers were randomly divided into three groups: Experimental Example 3-1-1, Experimental Example 3-1-2, and Experimental Example 3-1-3, with six participants in each group. The volunteers were aged 18-50 years, both men and women (analysis was stratified by gender). Participants were required to have mild to moderate hair loss, had not used hair growth medications in the past three months, and had no serious scalp conditions. Apply 1 mL of the extract evenly to the scalp once daily, massaging with the fingertips until absorbed. After the study, scalp photographs were taken using a trichoscope.

[0090] After using the test sample for 28 days, hair images of designated areas were collected using a hair mirror. Representative results of volunteers are shown below: Figure 7 As shown in the figure, each experimental example has an effect on the scalp microenvironment. The scalp microenvironment of the subjects was significantly improved, and erythema and abnormal oil secretion were significantly reduced. In addition, the dandruff of the subjects in experimental example 3-1-3 was significantly reduced.

[0091] In order to further explore the effect of Lactobacillus plantarum fermentation products on the scalp microenvironment, scalp essences containing different amounts of conditioners were prepared, and 0.1%, 1%, 5% and 10% of Lactobacillus plantarum fermentation products were added (Experimental Example 3-2-1, Experimental Example 3-2-2, Experimental Example 3-2-3 and Experimental Example 3-2-4), respectively. It also included 1% Ilex kudingcha leaf extract, 5% dimethyl isosorbide, 0.2% methylparaben, 0.35% phenoxyethanol, 0.8% ethylhexylglycerin, 0.3% D-panthenol, 5% butylene glycol, 0.2% betaine and 0.5% EDTA-2Na, and the balance was water.

[0092] Suitable volunteers were recruited for a hair loss prevention test using Kudingcha Wintergreen Extract. Twenty-four volunteers were randomly divided into four groups: Experimental Example 3-2-1, Experimental Example 3-2-2, Experimental Example 3-2-3, and Experimental Example 3-2-4, with six participants in each group. The volunteers were aged 18-50 years, both men and women (analysis stratified by gender). Participants were required to have mild to moderate hair loss, not used hair growth medications in the past three months, and not have severe scalp conditions. Apply 1 mL of the extract evenly to the scalp once daily and massage with the fingertips until absorbed. After the test, scalp photographs were taken using a trichoscope.

[0093] After using the test sample for 28 days, hair images of designated areas were collected using a hair mirror. Representative results of volunteers are shown below: Figure 8 As shown in the figure, Experimental Examples 3-2-1, 3-2-2, 3-2-3, and 3-2-4 all had significant effects on the scalp microenvironment, significantly improving the scalp microenvironment and alleviating erythema and abnormal oil secretion. Furthermore, the dandruff of the subject in Experimental Example 3-2-3 was significantly reduced. The overall effect was best in Experimental Example 3-2-3, followed by Experimental Example 3-2-4, which was superior to Experimental Examples 3-2-2 and 3-2-1.

[0094] Experimental Example 7: Product Ratio Study

[0095] According to the formula design of Kudingcha Ilex scalp essence, the core active substance Kudingcha Ilex scalp extract, conditioner and penetration enhancer were comprehensively formulated to evaluate the anti-hair loss efficacy under different formulations.

[0096] Table 6

[0097]

[0098] The mass percentage of Ilex kudingcha leaf extract added to the scalp essence of each group was 1%, and the addition ratios of conditioners and penetration enhancers were shown in Table 6. The scalp essence also included 0.2% methylparaben, 0.35% phenoxyethanol, 0.8% ethylhexylglycerin, 0.3% D-panthenol, 5% butylene glycol, 0.2% betaine and 0.5% EDTA-2Na, with the balance being water.

[0099] According to the evaluation method of male hair loss model mice in Example 4, 30 male C57BL / 6J mice were randomly divided into 6 groups: blank group, model group, experimental example 4-1, experimental example 4-2, experimental example 4-3, and experimental example 4-4. Except for the blank group, the other groups were evenly applied with 200 μL of testosterone (5 mg / mL) every day for modeling treatment, and 200 μL of the test substance was applied after complete absorption. The model group was treated with a blank matrix of essence without active substances for 14 consecutive days. After the experimental mice were anesthetized, the hair growth in the hair loss area on the back of the mice was photographed, and then a 1cm 2 A hair loss test was performed on the back test area using transparent tape. The number of hairs removed was counted to reflect the anti-hair loss effect of each group. Any hair that grew in the shaved area was shaved with a shaver, collected with weighing paper, and weighed using a 1 / 100,000 electronic balance.

[0100] The results showed that the effects of the four groups of samples on the amount of hair loss and hair weight in mice with androgenic alopecia were as follows: Figure 9 As shown in A, B, and C. The results show that Experimental Examples 4-1 through 4-4 all showed significant anti-hair loss effects, significantly inhibiting hair loss in male mice, reducing the number of hair loss episodes, and increasing hair weight in the affected areas. The treatment group in Experimental Example 4-3 (containing 3% Ilex kudingcha leaf extract, 5% conditioner, and 5% penetration enhancer) showed the best results. The overall anti-hair loss effectiveness of the four experimental examples was ranked as follows: Experimental Example 4-3, Experimental Example 4-4, Experimental Example 4-1, and Experimental Example 4-2. The mass ratio of Ilex kudingcha leaf extract, conditioner, and penetration enhancer was 1:2-7:4-10; preferably 1:5:8.

[0101] Experimental Example 8

[0102] A scalp essence comprising 1% Ilex kudingcha leaf extract, 5% Lactobacillus plantarum fermentation product, 5% dimethyl isosorbide, 0.2% methylparaben, 0.35% phenoxyethanol, 0.8% ethylhexylglycerin, 0.3% D-panthenol, 5% butylene glycol, 0.2% betaine, and 0.5% EDTA-2Na, with the balance being water, was prepared as Experimental Example 5-1; a scalp essence comprising 45% ethanol was also constructed as Experimental Example 5-2.

[0103] The scalp essence includes, by weight percentage, 1% Ilex kudingcha leaf extract, 5% Lactobacillus plantarum fermentation product, 5% dimethyl isosorbide, 0.3% caprylyl glycol, 0.3% 1,2-hexanediol, 0.2% p-hydroxyacetophenone, 2% propylene glycol, and 0.5% EDTA-2Na, with the balance being water, as Experimental Example 5-3.

[0104] Suitable volunteers were recruited for a hair loss prevention test using Kudingcha Wintergreen Essence. Subjects were randomly divided into 6 groups, aged 18-50 years, both men and women (stratified by gender). Participants were required to have mild to moderate hair loss, not used hair growth medications in the past 3 months, and no serious scalp diseases. Once daily, 1 mL was applied evenly to the scalp and massaged until absorbed. The group tests were conducted according to the essence preparation methods described in Experimental Examples 5-1-5-3. After 28 days of use, hair images of designated areas were collected using a trichoscope. The results for representative volunteers are shown below. Figure 10 As shown, Experimental Examples 5-1, 5-2, and 5-3 all had significant effects on hair loss. The subjects' scalp microenvironment was significantly improved, erythema and abnormal oil secretion were significantly reduced, a large number of new hairs grew, the overall hair density increased, and the hair roots were stronger than before use. In addition, the amount of hair loss was significantly reduced. The results are shown in Table 7. Among them, Experimental Example 5-2 had the best anti-hair loss effect, with a hair loss improvement rate of 78.5%; Experimental Example 5-1, a hair loss improvement rate of 64.2%; Experimental Example 5-3, a hair loss improvement rate of 53.3%. In summary, the Kudingcha Ilex Leaf Essence has a significant anti-hair loss effect.

[0105] Table 7 Statistics of hair loss

[0106]

[0107] The embodiments of the present invention are described in detail above, but the contents described are only preferred embodiments of the present invention and should not be considered to limit the scope of the present invention. All equivalent changes and improvements made within the scope of the present invention should still fall within the scope of the patent coverage of the present invention.

Claims

1. An anti-hair loss essence containing Ilex kudingcha leaf extract, characterized by: It includes Kudingcha Ilex leaf extract and lactobacillus fermentation product; Kudingcha Ilex leaf is extracted by decocting in water, the filtrate obtained by decoction is concentrated and dried, and the obtained solid substance is Kudingcha Ilex leaf extract. The Kudingcha Ilex leaf extract in the anti-hair loss essence accounts for 0.05%-10% by weight; the lactobacillus fermentation product is used as a conditioning agent, accounting for 0.1-10% by weight.

2. The anti-hair loss essence containing Ilex kudingcha leaf extract according to claim 1, characterized in that: The lactobacillus fermentation product is a lactobacillus fermentation lysate or a Lactobacillus plantarum fermentation product.

3. The anti-hair loss essence containing Ilex kudingcha leaf extract according to claim 1, characterized in that: The Lactobacillus fermentation product was replaced with Pichia pastoris fermentation lysate filtrate.

4. The anti-hair loss essence containing Ilex kudingcha leaves extract according to any one of claims 1 to 3, characterized in that: The invention also comprises 2-8% of a penetration enhancer, 0.8-1.35% of a preservative, 2-15.5% of a moisturizing agent, 0.1-1% of a chelating agent, and the balance being a solvent.

5. The anti-hair loss essence containing Ilex kudingcha leaves extract according to claim 4, characterized in that: The mass ratio of the Ilex kudingcha leaf extract, the conditioner and the penetration enhancer is 1:2-7:4-10.

6. The anti-hair loss essence containing Ilex kudingcha leaves extract according to claim 4, characterized in that: The penetration enhancer is dimethyl isosorbide and / or ethoxydiglycol.

7. The anti-hair loss essence containing Ilex kudingcha leaves extract according to claim 4, characterized in that: The preservative is selected from one or more of methylparaben, caprylyl glycol, phenoxyethanol, ethylhexylglycerin, 1,2-hexanediol, and p-hydroxyacetophenone; The moisturizing agent is selected from one or more of glycerin, D-panthenol, propylene glycol, butylene glycol, and betaine; The solvent is water or a mixture of water and ethanol.

8. A method for preparing the anti-hair loss essence containing the Ilex kudingcha leaf extract according to any one of claims 1 to 7, characterized in that: Add the solvent, Ilex kudingcha leaf extract, preservative, moisturizer, and chelating agent into the emulsification tank, start stirring at a speed of 20-40 rpm; heat to 80°C and keep warm for 10-15 minutes; The mixture is stirred and cooled to 40-50°C, a conditioner is added, and the mixture is stirred evenly until the temperature is below 40°C. The mixture is removed from the pot and left to stand for a while to obtain an anti-hair loss essence containing the Kudingcha Ilex leaf extract.

9. The method for preparing the anti-hair loss essence containing Ilex kudingcha leaves extract according to claim 8, characterized in that: Adjust the pH value of the anti-hair loss serum to 5.0-7.

0.

10. The method for preparing the anti-hair loss essence containing Ilex kudingcha leaves extract according to claim 8, characterized in that: The Kudingcha Ilex leaf extract includes Kudingsaponin active substances.

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