A non-removable hair growth composition and its use in hair growth products

Abstract

This invention relates to the field of hair growth composition technology, specifically to an anti-hair loss and hair growth composition and its application in hair growth products. The active ingredients of the composition include GHK-Gu and Erigeron tiglium extract, and preferably also include Eclipta prostrata extract. Through in-depth research, this invention has for the first time discovered that GHK-Gu and Erigeron tiglium extract can produce a synergistic effect at a specific ratio. Based on this, a ternary compound system containing Eclipta prostrata extract was optimized. The composition significantly improves hair density, diameter, and color through multi-target synergistic effects, including inhibiting 5α-reductase activity, promoting dermal papilla cell proliferation, and activating tyrosinase. This invention solves the technical problems of significant side effects from existing chemical drugs and complex compositions of traditional Chinese medicine compound formulas, providing a highly efficient, safe, and simplified solution for hair loss treatment. In vitro and in vivo experiments have confirmed its significant anti-hair loss, hair growth, and hair darkening effects.

Description

Technical Field

[0001] This invention relates to the field of hair growth composition technology, and in particular to an anti-hair loss and hair growth composition and its application in hair growth products. Background Technology

[0002] Hair loss is a common hair disorder with a complex pathological mechanism, mainly involving androgen-mediated miniaturization of hair follicles (5α-reductase converts testosterone to dihydrotestosterone, DHT), microinflammation and oxidative stress around hair follicles, decreased growth factor secretion due to the decline in dermal papilla cell function, and hair follicle cycle disorder (shortened anagen phase). Androgenetic alopecia (AGA) is the most common type clinically.

[0003] Currently, the mainstream clinical treatments include minoxidil and finasteride. Minoxidil promotes blood supply to hair follicles by dilating blood vessels, but it has problems such as local irritation, hirsutism, and relapse after discontinuation. Finasteride reduces DHT production by inhibiting 5α-reductase, but may be accompanied by side effects such as sexual dysfunction. Although topical Chinese medicine preparations have good safety, most existing formulations are complex, contain numerous ingredients, have unclear mechanisms of action, and lack rigorous verification by modern pharmacology. For example, although there are reports of combining Polygonum multiflorum, Ligustrum lucidum, and Platycladus orientalis leaves (such as CN119607093A), their composition is complex, and the synergistic effect mechanism is unclear.

[0004] In recent years, peptide-based active ingredients have shown potential in the field of hair growth. Chinese invention patent CN105106028B discloses a composition containing peptides that promote collagen production and peptides that promote keratin production, but this technology mainly focuses on simple combinations of peptides. Chinese invention patent CN110731922B discloses a combination of concentrated birch sap and other extracts, but its core lies in the concentration process of birch sap. Chinese invention patent CN118370806B discloses a compound hair growth composition containing privet fruit, fleeceflower root, perilla leaf, and various peptides and collagen; its components are numerous and its preparation process is complex.

[0005] GHK-Gu (a copper peptide derivative), a bioactive peptide with collagen-promoting, antioxidant, and anti-inflammatory activities, has been shown to stimulate hair follicle growth. However, its synergistic effects with other plant extracts have not been fully investigated. Given the shortcomings of existing technologies, developing a hair growth composition with simplified components, a clear mechanism of action, multi-target synergy, and high safety remains a pressing technical challenge in this field. Summary of the Invention

[0006] To address the aforementioned technical problems, this invention, through extensive experimental screening and verification, has for the first time discovered that GHK-Gu and Erigeron brevis extract can produce a significant synergistic effect at a specific ratio. Based on this, a ternary compound system containing Erigeron brevis extract was further optimized. This composition, through multi-target and multi-pathway synergistic effects, exhibits excellent results in inhibiting DHT production, promoting hair follicle microcirculation, anti-inflammatory and antioxidant effects, and promoting melanin production.

[0007] To achieve the above-mentioned technical effects, the present invention adopts the following technical solution: In a first aspect, the present invention provides a hair loss prevention and hair growth composition, the active ingredients of which, by mass parts, comprise the following raw material components: 0.5-10 parts of GHK-Gu and 5-15 parts of Erigeron tiglium extract.

[0008] In the technical solution of this invention, GHK-Gu (a copper peptide derivative), as a signal peptide, can effectively stimulate the proliferation of dermal papilla cells in hair follicles and upregulate the expression of vascular endothelial growth factor (VEGF), thereby improving the microcirculation around the hair follicles and providing sufficient nutrition for hair growth. Erigeron brevis extract is rich in flavonoids, caffeic acid esters, and other active ingredients, possessing significant antioxidant and anti-inflammatory effects. It can scavenge free radicals, protect hair follicle cells from oxidative stress damage, and inhibit inflammatory responses, creating a healthy environment for hair growth. The combined use of these two ingredients can synergistically enhance the effects of promoting hair follicle vitality and protecting the hair follicle microenvironment, achieving the goal of preventing hair loss and promoting hair growth.

[0009] As an optional technical solution, the number of GHK-Gu parts can be 0.5 parts, 1 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, or 10 parts, or any range between these values. The number of parts of the Erigeron brevis extract can be 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, or 15 parts, or any range between these values.

[0010] Secondly, the present invention provides a hair loss prevention and hair growth composition, wherein the active ingredients, in addition to the components described in the first aspect, include 1-10 parts by mass of Eclipta prostrata extract.

[0011] Eclipta prostrata extract is a traditional Chinese medicine for hair growth and darkening. Its main active ingredients, such as wedelia lactone, have antioxidant, anti-inflammatory, and immunomodulatory effects. It can also activate tyrosinase activity, promote melanin production, and help improve the color of new hair growth, making it black and shiny. Adding Eclipta prostrata extract to the first-part composition not only further enhances the antioxidant and anti-inflammatory protective effects but also additionally improves hair quality and darkens hair, making the product more comprehensive.

[0012] As an optional technical solution, the number of parts of the Eclipta prostrata extract can be 1 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts or 10 parts, and any range of values ​​between these points.

[0013] Thirdly, the present invention provides a hair loss prevention and hair growth composition, the active ingredients of which, by mass parts, comprise the following raw material components: 2-6 parts of GHK-Gu, 6-12 parts of Erigeron tiglium extract and 2-6 parts of Eclipta prostrata extract.

[0014] This formulation range represents a further optimization based on the aforementioned scheme, aiming to achieve the best synergistic effect of the three core active ingredients in a specific ratio. Within this range, the three can interact more efficiently, maximizing hair growth stimulation while comprehensively protecting hair follicles from internal and external factors, and simultaneously improving hair quality, thus achieving optimal hair growth and nourishment results.

[0015] As an optional technical solution, the number of GHK-Gu portions can be 2, 3, 4, 5, or 6, or any range between these point values. The number of Erigeron brevis extract portions can be 6, 7, 8, 9, 10, 11, or 12, or any range between these point values. The number of Eclipta prostrata extract portions can be 2, 3, 4, 5, or 6, or any range between these point values.

[0016] Fourthly, the present invention provides a hair loss prevention and hair growth composition, the active ingredients of which, by mass parts, comprise the following raw material components: 4 parts GHK-Gu, 8 parts Erigeron tiglium extract and 4 parts Eclipta prostrata extract.

[0017] This is the optimal formulation that has been experimentally verified to have the most outstanding effect. Under this specific formulation, the synergistic effect of GHK-Gu, Erigeron brevis extract, and Eclipta prostrata extract reaches its peak, exhibiting optimal levels in multiple functions such as inhibiting 5α-reductase activity, promoting dermal papilla cell proliferation, anti-oxidation, and activating tyrosinase.

[0018] Fifthly, the present invention provides a method for preparing a hair loss prevention and hair growth composition according to any one of the first to fourth aspects, comprising the following steps: S1: Preparation of extracts from Erigeron breviscapus and Eclipta prostrata, including: Take the dried whole plant of *Erigeron breviscapus*, pulverize it, and pass it through a 20-100 mesh sieve. Weigh a certain amount and place it in an extraction container. Add 10-30 times (v / w) of 50-95% ethanol solution, and extract by reflux at 60-80℃ 1-3 times, 1-2 hours each time. Filter the extract, combine the filtrates, and concentrate under reduced pressure at 50-60℃ to recover the ethanol, obtaining a concentrated solution. Further dry the concentrated solution, either by freeze-drying or spray-drying, to obtain *Erigeron breviscapus* extract powder. Take the dried whole herb of Eclipta prostrata, pulverize it, and pass it through a 20-100 mesh sieve. Weigh a certain amount and place it in an extraction container. Add 10-30 times (v / w) of 50-95% ethanol solution, and extract by reflux at 60-80℃ 1-3 times, 1-2 hours each time. Filter the extract, combine the filtrates, and concentrate under reduced pressure at 50-60℃ to recover the ethanol, obtaining a concentrated solution. Further dry the concentrated solution to obtain Eclipta prostrata extract powder. S2: Weigh out GHK-Gu, Erigeron brevis extract and Eclipta prostrata extract according to the formula, mix them, and prepare them with suitable excipients to make a topical hair growth product.

[0019] This preparation method is simple to operate and operates under mild conditions, which can extract and retain the effective active ingredients in Erigeron breviscapus and Eclipta prostrata to the maximum extent, ensuring the high quality and bioactivity of the raw materials for the composition.

[0020] As an optional technical solution, in step S1, the extraction conditions of the extracts of Erigeron breviscapus and Eclipta prostrata can be further optimized as follows: using 60-80% ethanol solution, a material-to-liquid ratio of 1:15-25, an extraction temperature of 70-80℃, extraction twice, and extraction for 1.5 hours each time.

[0021] As an optional technical solution, in step S2, the total concentration of active ingredients in the topical hair growth product can be between 0.1 wt% and 8 wt%. This concentration range ensures that the product has good hair growth and anti-hair loss effects while also ensuring its safety and skin tolerance during use. Too low a concentration may result in poor effects, while too high a concentration may increase the risk of irritation or cause unnecessary waste of raw materials.

[0022] As an optional technical solution, the topical hair growth product can be in various forms commonly found in the art, such as, but not limited to: hair growth liquid, shampoo, gel, serum, conditioner, scalp care mask, etc.

[0023] Sixthly, the present invention provides the use of the anti-hair loss and hair growth composition according to any one of the first to fourth aspects in the preparation of a hair growth product, said hair growth product having at least one of the following functions: (a) Inhibits 5α-reductase activity, reducing dihydrotestosterone production; (b) Promotes the proliferation of dermal papilla cells and the expression of vascular endothelial growth factor; (c) Activates tyrosinase, promotes melanin production in hair, and improves hair color; (d) Promotes the transformation of hair follicles from the resting phase to the growth phase, increasing hair density and diameter.

[0024] This composition intervenes in hair loss problems at their root through a multi-target, multi-pathway mechanism of action.

[0025] This invention is the first to discover a synergistic effect between GHK-Gu and Erigeron brevis extract in promoting hair growth. GHK-Gu effectively promotes the proliferation of dermal papilla cells and improves the hair follicle microenvironment; Erigeron brevis extract, on the other hand, can reduce the production of dihydrotestosterone at its source by inhibiting 5α-reductase activity and combat inflammation and oxidative stress around the hair follicle. The combined application of the two complements each other in promoting hair follicle vitality and blocking hair loss signals, achieving effective intervention in the pathological process of hair loss.

[0026] Building upon this foundation, the present invention further incorporates Eclipta prostrata extract, constructing a ternary compound system of GHK-Gu, Erigeron brevis extract, and Eclipta prostrata extract. Eclipta prostrata extract can activate tyrosinase, promote melanin synthesis, and help improve the color of newly grown hair. The synergistic effect of the three components demonstrates superior performance compared to single components or binary combinations in promoting dermal papilla cell proliferation, inhibiting 5α-reductase activity, and activating tyrosinase, achieving a dual improvement in both the quantity and quality of hair.

[0027] Compared with existing chemical drugs, the composition of this invention eliminates ingredients such as minoxidil and finasteride, and is entirely composed of bioactive peptides and natural plant extracts. Its mechanism of action is mild, avoiding local irritation and systemic side effects, resulting in higher safety. Compared with existing traditional Chinese medicine formulas, the composition of this invention has simplified components and clear proportions, overcoming the shortcomings of traditional Chinese medicine formulas such as complex ingredients and unclear mechanisms of action. The product has stable efficacy, controllable quality, and is easier to industrialize.

[0028] The composition of this invention can more comprehensively intervene in the complex pathological process of hair loss through the synergistic effect of multiple targets and pathways, and shows superior application potential in the treatment of various types of hair loss such as androgenetic alopecia. Detailed Implementation

[0029] The following embodiments are only used to more clearly illustrate the technical solutions of the present invention. The specific embodiments listed in the present invention are only examples of the present invention, and the present invention is not limited to the specific embodiments described below.

[0030] Unless otherwise specified, all materials used in the following examples are commercially available. GHK-Gu (CAS No.: 130120-56-2) was purchased from Chengdu Shengnuo Biopeptide Technology Co., Ltd., Erigeron acris L. (family Asteraceae) is the whole herb, and Eclipta prostrata L. (family Asteraceae) is the whole herb.

[0031] Example 1

[0032] The purpose of this embodiment is to provide extracts of Eclipta prostrata and Eclipta prostrata, and their preparation methods, as follows: (1) Preparation of Erigeron brevis extract: Dried whole *Erigeron tigrinum* herb was pulverized and passed through a 40-mesh sieve. 12 times the volume (v / w) of 70% ethanol was added, and the mixture was extracted twice under reflux at 75°C, 1.5 hours each time. The two extracts were combined, filtered, and the filtrate was concentrated under reduced pressure at 55°C until no alcohol odor remained, yielding an extract. The extract was dispersed in an appropriate amount of water and freeze-dried to obtain *Erigeron tigrinum* extract powder, with a yield of approximately 8.5%.

[0033] (2) Preparation of Eclipta prostrata extract: The dried whole herb of Eclipta prostrata was pulverized and passed through a 40-mesh sieve. Ten times the volume (v / w) of 60% ethanol was added, and the mixture was extracted three times under reflux at 70°C, each time for 2 hours. The extracts were combined, filtered, and the filtrate was concentrated under reduced pressure at 60°C until no alcohol odor remained, yielding an extract. The extract was dispersed in an appropriate amount of water and freeze-dried to obtain Eclipta prostrata extract powder, with a yield of approximately 10.2%.

[0034] Example 2

[0035] To screen for the optimal ratio, a series of composition samples were prepared according to the ratios shown in Table 1 in this embodiment. The specific procedures are as follows: Accurately weigh GHK-Gu and the extracts of *Erigeron breviscapus* and *Eclipta prostrata* prepared in Example 1, and mix them according to the corresponding mass fractions in Table 1 (the total mass fraction of each sample is 10 parts, but the mass fractions only represent relative proportions; the actual preparation should be scaled up proportionally). Place each weighed component in a beaker, add an appropriate amount of deionized water, and stir magnetically until completely dissolved. Transfer to a volumetric flask, add deionized water to the mark, and prepare a test sample solution with a total active ingredient concentration of 1.5 wt% (GHK-Gu, *Erigeron breviscapus* extract, and *Eclipta prostrata* extract are all considered as active ingredients). Prepare 100 mL of each sample for later use. All samples should be prepared fresh or stored at 4°C (for no more than one week).

[0036] Table 1 lists the specific formulations of the 13 samples used in subsequent pharmacodynamic experiments. Samples 1 to 3 are binary combinations of GHK-Gu and Erigeron brevis extract, examining the effect of different GHK-Gu dosages; samples 4 to 8 are ternary combinations of GHK-Gu, Erigeron brevis extract, and Eclipta prostrata extract, examining variations in the proportions of different components; sample 5 represents the preferred ternary optimal formulation of this invention. Samples 9 to 13 are comparative examples, including the GHK-Gu single-use group (sample 9), the Erigeron brevis extract single-use group (sample 10), the Eclipta prostrata extract single-use group (sample 11), and the binary combination of GHK-Gu and Eclipta prostrata extract (sample 12) and the binary combination of Erigeron brevis extract and Eclipta prostrata extract (sample 13), used for comparative analysis in subsequent experiments.

[0037] Table 1. Samples of compositions with different proportions (parts by mass) Sample number GHK-GU (portions) Erigeron brevis extract (per serving) Eclipta prostrata extract (per serving) Sample 1 2 8 0 Sample 2 4 8 0 Sample 3 6 8 0 Sample 4 4 4 4 Sample 5 4 5 4 Sample 6 4 12 4 Sample 7 2 8 4 Sample 8 4 8 2 Sample 9 4 0 0 Sample 10 0 8 0 Sample 11 0 0 4 Sample 12 4 0 4 Sample 13 0 8 4 Experimental Example 1 To investigate the mechanism of action of the composition of the present invention in preventing hair loss and promoting hair growth at the cellular and molecular levels, the following in vitro pharmacodynamic studies were conducted.

[0038] 1.1 Hair papilla cell proliferation experiment Experimental Methods: Rat dermal papilla cells (DPCs) from passages 3-5 were routinely cultured in DMEM medium containing 10% fetal bovine serum. When cells reached 80% confluence, they were digested with 0.25% trypsin and seeded at a density of 5 × 10³ cells per well (100 μL per well). The cells were incubated at 37°C with 5% CO2 for 24 hours to allow adhesion. The original medium was discarded, and 100 μL of medium containing 1.5% of the test sample (samples 1-13) was added to each well (6 replicates per sample). A blank control group (without sample) was also included. After 48 hours of further culture, 10 μL of CCK-8 solution was added to each well, and the cells were incubated for 2 hours. The absorbance (OD) at 450 nm was measured using a microplate reader. The relative cell proliferation rate was calculated using the following method: Relative proliferation rate (%) = (mean OD of experimental group / mean OD of blank control group) × 100%.

[0039] The relative cell proliferation rates of each group are shown in Table 2.

[0040] Table 2 Results of dermal papilla cell proliferation experiment (X±s, n=6) Sample number Relative proliferation rate (%) Sample 1 138.5 ± 5.2* Sample 2 152.1 ± 7.1* Sample 3 145.7 ± 4.8* Sample 4 148.3 ± 5.5* Sample 5 171.6 ± 6.8*# Sample 6 160.2 ± 5.6*# Sample 7 155.8 ± 6.9* Sample 8 153.4 ± 5.6* Sample 9 118.6 ± 4.5 Sample 10 112.4 ± 4.2 Sample 11 105.8 ± 3.1 Sample 12 135.2 ± 4.9* Sample 13 128.9 ± 4.8* Note: * indicates p < 0.05 compared to sample 9; # indicates p < 0.05 compared to sample 2.

[0041] The results of the dermal papilla cell proliferation experiment showed significant differences in the cell proliferation-promoting effects of different samples. Both GHK-Gu and Erigeron brevis extract alone showed some proliferative effects, but the binary combination of the two was more potent, confirming a synergistic proliferative effect between GHK-Gu and Erigeron brevis extract. Sample 2, with a 4:8 ratio of GHK-Gu to Erigeron brevis extract, exhibited the highest proliferation rate among the binary groups. Eclipta prostrata extract alone showed the weakest proliferative effect, with the lowest proliferation rate among all tested samples.

[0042] When Eclipta prostrata extract was combined with GHK-Gu and Erigeron brevis extract, sample 5 showed the highest proliferation rate, significantly higher than both the GHK-Gu monotherapy group and sample 2, indicating that the ternary combination system further enhanced the proliferation-promoting effect. The proliferation rates of other ternary groups were generally higher than those of the binary groups, with sample 6 showing a significantly higher proliferation rate than sample 2, while samples 7 and 8, although better than sample 2, did not reach statistical significance. Overall, the ternary combination showed superior performance in promoting dermal papilla cell proliferation compared to the binary combination and the monotherapy groups, with sample 5 exhibiting the most outstanding effect.

[0043] 1.2 5α-Reductase Inhibition Experiment Experimental Methods: Crude 5α-reductase from rat testes was prepared according to the literature method. Healthy male SD rats were sacrificed, and their testes were quickly removed, washed in pre-cooled physiological saline, the membrane was removed, and the testes were weighed. Four volumes of 0.1 mol / L Tris-HCl buffer were added, and the mixture was homogenized in an ice bath. The homogenate was centrifuged at 12,000 rpm for 20 minutes at 4°C, and the supernatant was used as the crude enzyme solution. Protein concentration was determined by the BCA method. The total reaction volume was 200 μL, containing: 0.1 mol / L Tris-HCl buffer (pH 7.4), 0.5 mmol / L NADPH, 10 μmol / L testosterone, an appropriate amount of crude enzyme solution (approximately 100 μg protein), and different test samples (final concentration 50 μg / mL). A blank control (without sample) was also included. After incubation at 37°C for 30 minutes, 200 μL of ice-cold acetonitrile was immediately added to terminate the reaction. The supernatant was collected by centrifugation, and the remaining testosterone content was detected using a testosterone ELISA kit. Calculate the 5α-reductase inhibition rate: Inhibition rate (%) = (1 - testosterone reduction in the sample group / testosterone reduction in the blank control group) × 100%.

[0044] The inhibition rates of 5α-reductase in each group are shown in Table 3.

[0045] Table 3. Results of 5α-reductase inhibition experiment (X±s, n=3) Sample number Inhibition rate (%) Sample 1 62.3 ± 4.2* Sample 2 71.5 ± 3.4* Sample 3 67.8 ± 2.5* Sample 4 69.2 ± 3.3* Sample 5 84.7 ± 5.0*# Sample 6 78.9 ± 4.4*# Sample 7 73.6 ± 4.1* Sample 8 72.1 ± 3.3* Sample 9 45.8 ± 3.4 Sample 10 55.3 ± 3.5* Sample 11 18.5 ± 1.9 Sample 12 52.4 ± 3.1 Sample 13 60.7 ± 4.2* Note: * indicates p < 0.05 compared to sample 9; # indicates p < 0.05 compared to sample 2.

[0046] The 5α-reductase inhibition assay showed significant differences in inhibitory activity among different samples. Erigeron brevis extract alone exhibited moderate inhibitory activity, while GHK-Gu alone showed weak inhibitory activity. Eclipta prostrata extract alone showed the lowest inhibition rate, being the least effective among all tested samples. The combined use of GHK-Gu and Erigeron brevis extract significantly improved the inhibition rate, with sample 2 showing a significantly higher inhibition rate than the GHK-Gu-Gu-alone group, confirming a synergistic effect between the two in inhibiting 5α-reductase.

[0047] When Eclipta prostrata extract was added to form a ternary system, sample 5 showed the highest inhibition rate, significantly higher than both the GHK-Gu monotherapy group and sample 2, indicating that the addition of Eclipta prostrata extract further enhanced the combination's ability to block the androgen pathway. Among the other ternary groups, sample 6 also showed a significantly higher inhibition rate than sample 2, while samples 7 and 8 showed no significant difference compared to sample 2, suggesting that the ratio of GHK-Gu to Eclipta prostrata extract has a significant impact on the synergistic effect. Overall, the ternary compound system exhibited better 5α-reductase inhibitory activity than the binary combination and the single components, with sample 5 showing the most outstanding effect.

[0048] 1.3 Experiment on Tyrosinase Activity and Melanin Production Experimental method: B16F10 mouse melanoma cells in logarithmic growth phase were used, with 1×10⁶ cells per well. 5 Cells were seeded at a density of 2 mL of culture medium in each well of a 6-well plate. After 24 hours of culture, the original culture medium was discarded, and culture medium containing 1.5% of the test sample was added to each well, with three replicates for each sample. A blank control group was also included. After 48 hours of further culture, the supernatant was discarded, and the cells were washed twice with PBS. 200 μL of cell lysis buffer (PBS containing 1% Triton X-100) was added to each well, and the cells were lysed on ice for 30 minutes. Cells were scraped off, and the lysis buffer was collected. The cells were centrifuged at 12,000 rpm for 10 minutes at 4°C. The supernatant was used for tyrosinase activity assay and protein quantification. Tyrosinase activity assay: 80 μL of the supernatant was added to a 96-well plate, and 20 μL of 10 mmol / L levodopa (L-DOPA) was added. The plate was incubated at 37°C for 30 minutes, and the absorbance at 490 nm was measured using a microplate reader.

[0049] Melanin content determination: A separate sample of cells was digested with trypsin, counted, and centrifuged to collect the cell pellet. 1 mL of 1 mol / L NaOH solution containing 10% DMSO was added, and the mixture was heated at 80°C for 1 hour to dissolve the melanin. The absorbance at 405 nm was measured, and the melanin content was calculated based on the standard curve. Results are expressed as a percentage relative to the blank control group.

[0050] The effects of each group on tyrosinase activity and melanin content are shown in Table 4.

[0051] Table 4. Experimental results of tyrosinase activity and melanin production (X±s, n=3) Sample number Relative tyrosinase activity (%) Relative melanin content (%) Sample 1 125.6 ± 4.8 133.2 ± 5.5 Sample 2 138.4 ± 5.2 147.8 ± 7.1* Sample 3 133.1 ± 5.0 141.5 ± 4.8 Sample 4 155.3 ± 5.9* 168.2 ± 6.4* Sample 5 182.5 ± 7.0*# 205.4 ± 7.2*# Sample 6 169.8 ± 6.5* 185.3 ± 6.3*# Sample 7 162.4 ± 6.2 176.9 ± 7.1* Sample 8 166.7 ± 6.1* 180.5 ± 6.2* Sample 9 108.5 ± 4.3 112.3 ± 4.6 Sample 10 105.1 ± 4.2 109.5 ± 4.4 Sample 11 148.6 ± 5.5 162.7 ± 5.3 Sample 12 138.2 ± 5.3 151.6 ± 6.6 Sample 13 142.5 ± 5.5 158.3 ± 4.2 Note: * indicates p < 0.05 compared to sample 11; # indicates p < 0.05 compared to sample 2.

[0052] As shown in Table 4, the results of the tyrosinase activity and melanin production experiment indicate that Eclipta prostrata extract alone significantly promotes tyrosinase activity and melanin production, while GHK-Gu and Erigeron brevis extract alone have weaker effects and no significant promoting effect on melanin synthesis. In the binary combinations, the combination containing only Eclipta prostrata extract with GHK-Gu or Erigeron brevis extract has a certain increase in tyrosinase activity and melanin content, but the effect is not as significant as that of the group using Eclipta prostrata extract alone.

[0053] When GHK-Gu, Erigeron brevis extract, and Eclipta prostrata extract were combined, sample 5 showed the highest tyrosinase activity and melanin content, significantly higher than the group using Eclipta prostrata extract alone. This indicates that the addition of GHK-Gu and Erigeron brevis extract can synergistically enhance the effect of Eclipta prostrata extract, jointly promoting melanin synthesis. Based on the results of the three in vitro pharmacodynamic experiments, GHK-Gu and Erigeron brevis extract in the composition of this invention, at a specific ratio, synergistically promote dermal papilla cell proliferation and inhibit 5α-reductase, while the addition of Eclipta prostrata extract further synergizes with both, significantly enhancing tyrosinase activity and melanin production. Among them, sample 5 (GHK-Gu: Erigeron brevis extract: Eclipta prostrata extract = 4:8:4) showed the best performance in all indicators and is the preferred ratio of this invention.

[0054] Experimental Example 2

[0055] To verify the efficacy of the composition of the present invention in preventing hair loss and promoting hair growth at the in vivo level, an in vivo pharmacodynamic study was conducted using a testosterone-induced C57BL / 6 mouse androgenic alopecia model.

[0056] 1. Experimental Methods 1.1 Animal grouping and model establishment Sixty male C57BL / 6 mice aged 6-8 weeks were selected and, after one week of acclimatization, were randomly divided into 6 groups of 10 mice each. Blank control group: No treatment was given to the skin on the back.

[0057] Model control group: Subcutaneous injection of testosterone in the back + daily application of solvent (75% ethanol solution) control.

[0058] Positive control group: Subcutaneous injection of testosterone in the back + daily application of 5% minoxidil solution.

[0059] Sample 5 low-dose group: subcutaneous injection of testosterone in the back + daily application of a solution containing 0.75% active ingredient (sample 5).

[0060] Sample 5 medium dose group: subcutaneous injection of testosterone in the back + daily application of a solution containing 1.5% active ingredient (sample 5).

[0061] Sample 5 high-dose group: subcutaneous injection of testosterone in the back + daily application of a solution containing 3.0% active ingredient (sample 5).

[0062] Except for the blank control group, all other groups of mice were subcutaneously injected with testosterone solution (5 mg / kg body weight, dissolved in soybean oil) once daily for 21 consecutive days to establish an AGA model. From the first day of modeling, each group applied the corresponding test substance to the same area 4 hours after the testosterone injection once daily for 21 consecutive days.

[0063] 1.2 Experimental Results and Analysis Macroscopic hair regeneration: On day 21 after drug administration, photographs were taken of the skin on the backs of mice to observe changes in skin color (from pink to grayish-black, indicating that hair follicles have entered the growth phase) and the coverage of newly grown hair. The results showed that the skin on the backs of mice in the model control group remained pink throughout the experimental period, with only a very small amount of fine vellus hair growth, indicating that the androgenetic alopecia model was successfully established. Mice in the blank control group had dense hair on their backs. Compared with the model control group, all drug-treated groups showed varying degrees of hair regeneration promotion.

[0064] Hair density and diameter determination: On day 21, new hair was carefully shaved from the same area on the back of each mouse using a razor. 100 hairs were randomly selected from each group, and the hair diameter was measured under a microscope. At the same time, hair was collected from a fixed location on the back (1cm×1cm), the number of hair shafts was counted, and the hair density (hair / cm²) was calculated. The hair density and diameter measurement results of each group of mice are shown in Table 5.

[0065] Histopathological analysis: Mice were sacrificed on day 21, and skin from the dorsal drug administration area was collected, fixed in 10% formalin, embedded in paraffin, sectioned, and stained with hematoxylin and eosin (HE). The number of hair follicles in the anagen phase per field of view (100×) was observed and counted under an optical microscope. The experimental results are shown in Table 5.

[0066] Table 5 Effects on hair density and diameter in mice (X±s, n=10) Group Hair density (roots / cm²) Hair diameter (μm) Number of hair follicles in the growth phase (per field of view) Blank control group 285.6 ± 25.3 32.5 ± 3.1 18.5 ± 2.3 Model control group 102.3 ± 8.8# 18.7 ± 1.8# 5.2 ± 1.1# Positive control group (5% minoxidil) 238.5 ± 18.1* 28.4 ± 2.5* 14.8 ± 1.9* Sample 5, low-dose group (0.75%) 168.4 ± 16.5* 22.3 ± 1.7* 10.6 ± 1.6* Medium dose group (1.5%) in sample 5 205.7 ± 15.3* 26.1 ± 2.8* 12.3 ± 1.7* Sample 5, high-dose group (3.0%) 246.2 ± 19.6* 30.2 ± 2.9* 16.1 ± 1.9* Note: # indicates p < 0.05 compared with the blank control group; * indicates p < 0.05 compared with the model control group.

[0067] Experimental results showed that, compared with the model control group, all dose groups of Sample 5 significantly increased hair density and diameter in mice (p<0.05), and this effect was dose-dependent. The high-dose group of Sample 5 showed an effect close to that of the 5% minoxidil positive control group, and even slightly exceeded it in hair density, demonstrating excellent hair regrowth potential. Simultaneously, HE staining results showed a significant increase in the number of hair follicles in the anagen phase in the skin of mice in all dose groups of Sample 5, with increased follicle volume and the hair bulb penetrating deeper into the subcutaneous fat layer. This indicates that the composition of the present invention can effectively promote the transformation of hair follicles from the telogen phase to the anagen phase.

[0068] Example 3

[0069] The purpose of this embodiment is to provide a hair growth solution containing the preferred composition of the present invention and its preparation method, the formula of which (by mass parts) is as follows: The active ingredients (composed of GHK-Gu: Erigeron breviscapus extract and Eclipta prostrata extract in a mass ratio of 4:8:4) are 5 parts, ethanol (95%) is 50 parts, propylene glycol is 10 parts, 1,3-butanediol is 5 parts, menthol is 0.1 parts, and azone is 1.0 part. 0.05 parts sodium hyaluronate, 0.5 parts 1,2-hexanediol, and deionized water (to bring the total mass of the system to 100 parts). This hair growth serum is prepared using the following method: (1) Disperse sodium hyaluronate in a small amount of deionized water and swell at 40°C for 30 minutes to prepare a sodium hyaluronate solution.

[0070] (2) Mix the active ingredient with propylene glycol and 1,3-butanediol and stir until completely dissolved to obtain a viscous premix.

[0071] (3) Mix ethanol with azone and menthol, stir to dissolve, add the above viscous premixed liquid, stir evenly to obtain the ethanol phase.

[0072] (4) While stirring, slowly add the ethanol phase to the sodium hyaluronate solution, then add the remaining deionized water and 1,2-hexanediol, and continue stirring for 30 minutes until the system is uniform and transparent.

[0073] (5) Let stand to remove bubbles, filter, and fill to obtain the product.

[0074] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A hair loss prevention and hair growth composition, characterized in that, Its active ingredients, by mass, include the following raw material components: 0.5-10 parts of GHK-Gu and 5-15 parts of Erigeron brevis extract.

2. The hair loss prevention and hair growth composition as described in claim 1, characterized in that, The composition also includes 1-10 parts of Eclipta prostrata extract by weight of its active ingredients.

3. The hair loss prevention and hair growth composition as described in claim 2, characterized in that, Its active ingredients, by mass parts, include the following raw material components: 2-6 parts of GHK-Gu, 6-12 parts of Erigeron brevis extract, and 2-6 parts of Eclipta prostrata extract.

4. The hair loss prevention and hair growth composition as described in claim 3, characterized in that, Its active ingredients, by mass, include the following raw material components: 4 parts GHK-Gu, 8 parts Erigeron brevis extract, and 4 parts Eclipta prostrata extract.

5. A method for preparing a hair loss prevention and hair growth composition according to any one of claims 1-4, characterized in that, Includes the following steps: S1: Preparation of extracts from Erigeron breviscapus and Eclipta prostrata, including: Take the dried whole plant of *Erigeron breviscapus*, pulverize it, and pass it through a 20-100 mesh sieve. Weigh a certain amount and place it in an extraction container. Add 10-30 times (v / w) of 50-95% ethanol solution, and extract by reflux at 60-80℃ 1-3 times, 1-2 hours each time. Filter the extract, combine the filtrates, and concentrate under reduced pressure at 50-60℃ to recover the ethanol, obtaining a concentrated solution. Further dry the concentrated solution, either by freeze-drying or spray-drying, to obtain *Erigeron breviscapus* extract powder. Take the dried whole herb of Eclipta prostrata, pulverize it, and pass it through a 20-100 mesh sieve. Weigh a certain amount and place it in an extraction container. Add 10-30 times (v / w) of 50-95% ethanol solution, and extract by reflux at 60-80℃ 1-3 times, 1-2 hours each time. Filter the extract, combine the filtrates, and concentrate under reduced pressure at 50-60℃ to recover the ethanol, obtaining a concentrated solution. Further dry the concentrated solution to obtain Eclipta prostrata extract powder. S2: Weigh out GHK-Gu, Erigeron brevis extract and Eclipta prostrata extract according to the formula, mix them, and prepare them with suitable excipients to make a topical hair growth product.

6. The preparation method according to claim 5, characterized in that: The extraction conditions for the extracts of Erigeron breviscapus and Eclipta prostrata are as follows: using 50-95% ethanol solution, a material-to-liquid ratio of 1:10-30, an extraction temperature of 60-80℃, extraction 1-3 times, and extraction for 1-2 hours each time.

7. The preparation method according to claim 5, characterized in that, The total concentration of active ingredients in the topical hair growth product is 0.1 wt%-8 wt%.

8. The use of the hair growth promoting composition according to any one of claims 1-4 in the preparation of a topical hair growth product, wherein the hair growth product has at least one of the following functions: (a) Inhibits 5α-reductase activity, reducing dihydrotestosterone production; (b) Promotes the proliferation of dermal papilla cells and the expression of vascular endothelial growth factor; (c) Activates tyrosinase, promotes melanin production in hair, and improves hair color; (d) Promotes the transformation of hair follicles from the resting phase to the growth phase, increasing hair density and diameter.

9. The application as described in claim 8, characterized in that: The topical hair growth product is any one of the following: hair growth liquid, shampoo, gel, serum, conditioner, and scalp care mask.