Polypeptide-containing composition and application thereof in preparation of anti-alopecia products

Abstract

The invention belongs to the field of polypeptide compositions, and discloses a polypeptide-containing composition and an application thereof in preparation of an anti-alopecia product, the composition comprises: 40-100 parts of a Chinese patent medicine active matter, which is prepared by water extraction and alcohol precipitation of fleece-flower root, cacumen biotae, glossy privet fruit and yerbadetajo herb; 15-50 parts of plant active matter which contains ginseng fermentation liquor and ganoderan; the polypeptide compound is prepared from copper peptide GHK-Cu, palmitoyl tripeptide-1 and acetyl tetrapeptide-3 according to a specific proportion; the natural carrier is prepared from plant glycerin, camellia oil and seaweed gel. Through polypeptide synergistic interaction, natural carrier transdermal promotion and green process design, the problems of high chemical dependence, insufficient naturalness and unsustainable process in the prior art are solved, and the preparation method is suitable for preparing efficient and safe anti-alopecia products.

Description

Technical Field

[0001] The present invention relates to the technical field of polypeptide compositions, and in particular to a composition containing polypeptides and application thereof in preparing an anti-hair loss product. Background Art

[0002] The field of hair loss treatment has long faced a dual dilemma between chemically synthesized drugs and natural ingredient preparations. In the existing technology, although chemical drugs represented by minoxidil have certain therapeutic effects, they generally have the problem of high irritation and significant side effects. For example, the propylene glycol content in the minoxidil composition disclosed in the prior art is as high as 30%-50%. Long-term use can easily cause adverse reactions such as scalp dryness and contact dermatitis. Clinical studies have shown that the incidence of adverse reactions can reach 28.6%1. In addition, minoxidil relies on chemical solvents (such as ethanol) as a carrier, further exacerbating the risk of scalp barrier damage. Although some have tried to alleviate irritation by adding diosmin (a flavonoid compound), its formula is still mainly chemical ingredients and has failed to fundamentally solve the safety problem. More importantly, the preparation process of such compositions often involves extraction and emulsification treatment with toxic organic solvents (such as methanol), resulting in excessive emissions of volatile organic compounds (VOCs), which is contrary to the principles of green production.

[0003] On the other hand, although natural anti-hair loss compositions claim to be "chemical-free", they still have the defects of crude compatibility and inefficient process. For example, the plant extract composition disclosed in patent CN118045020A, although based on Chinese herbal medicines such as Polygonum multiflorum and Ligusticum chuanxiong, is added with paraben preservatives and chemical excipients such as propylene glycol. Accelerated tests show that the degradation rate of its active ingredients (such as flavonoids) during storage is as high as 42.3%. Such formulas not only fail to achieve true "natural and non-irritating", but also increase the risk of allergies due to the presence of chemical preservatives. In addition, traditional extraction processes mostly use high temperature and high pressure or organic solvent reflux methods. The extraction of Ganoderma lucidum polysaccharides needs to be carried out at 120°C, resulting in a loss rate of more than 60% of heat-sensitive components (such as adenosine), and the energy consumption intensity reaches 8.9kW·h / kg, which seriously deviates from the low-carbon and environmental protection goals. More prominently, the transdermal absorption rate of active ingredients in existing natural formulas is generally low. For example, in some cases, ginsenosides are combined with copper peptides. Due to the lack of a suitable carrier system, the transdermal efficiency is only 12.4±2.1%, and the two are prone to complex precipitation when pH>6.5, which significantly reduces bioavailability.

[0004] In summary, the main defects of existing technologies can be summarized as follows: high chemical dependence: mainstream formulas rely on chemical solvents such as propylene glycol and ethanol, which cause skin irritation after long-term use and a high recurrence rate after discontinuation of the drug; insufficient naturalness: compositions claiming to be "natural" still contain chemical preservatives or synthetic excipients, leading to doubts about safety and environmental protection; unsustainable process: the extraction process is energy-intensive and contains organic solvent residues (such as methanol residues >500ppm), which violates the principles of green chemistry; inefficient compatibility: the lack of a synergistic mechanism for active ingredients, low transdermal absorption rate, and difficulty in achieving stable therapeutic effects. These problems have severely limited the clinical application and market promotion of anti-hair loss products, and there is an urgent need to develop a truly green, efficient, and safe solution. Summary of the Invention

[0005] In order to solve the above-mentioned problems in the prior art, the present invention discloses a composition containing a polypeptide and its application in the preparation of an anti-hair loss product.

[0006] In order to achieve the above-mentioned purpose, the present invention solves the technical problem by adopting the following technical solutions:

[0007] In a first aspect, the present invention discloses a natural anti-hair loss composition, comprising the following components in parts by weight:

[0008] Active ingredient of Chinese patent medicine: 40-100 parts, prepared by mixing Polygonum multiflorum, Platycladus orientalis leaves, Ligustrum lucidum fruits, and Eclipta prostrata in a weight ratio of (5-15):(8-20):(3-12):(4-10), followed by water extraction and alcohol precipitation;

[0009] Plant active ingredients: 15-50 parts, including ginseng fermentation liquid and Ganoderma lucidum polysaccharide;

[0010] Peptide complex: 1-10 parts, composed of copper peptide GHK-Cu, palmitoyl tripeptide-1, and acetyl tetrapeptide-3 in a weight ratio of (1:0.5-2:0.3-1);

[0011] Natural carrier: 50-150 parts, including vegetable glycerin, camellia oil and algae gum.

[0012] Furthermore, in the above-mentioned natural anti-hair loss composition, the preparation method of the active ingredient of the Chinese patent medicine is:

[0013] The Chinese patent medicine forms of Polygonum multiflorum, Platycladus orientalis leaves, Ligustrum lucidum fruits and Eclipta prostrata are mixed in proportion, 5-15 times the weight of water are added and boiled for extraction 2-4 times, each time for 1-2 hours, and the combined extracts are concentrated to a relative density of 1.15-1.20, which is measured at 60°C, and precipitated with alcohol to a final concentration of 60-65% ethanol.

[0014] Furthermore, in the above-mentioned natural anti-hair loss composition, the plant active ingredients include:

[0015] The weight ratio of ginseng fermentation liquid to ganoderma lucidum polysaccharide is (2-3):1, and the preparation method of the ginseng fermentation liquid comprises the following steps:

[0016] (a) Wash and slice fresh ginseng, then mix with sterile water at a ratio of 1:3-5 (w / v);

[0017] (b) adding 3-5% sucrose solution and adjusting the sugar content of the final solution to 8-12°Bx;

[0018] (c) Inoculate Lactobacillus plantarum to a final concentration of 1×10 6 CFU / mL, fermented at 37-40°C for 72-96 hours;

[0019] (d) The supernatant was collected by centrifugation at 3000-5000 g, filtered through a 0.45 μm membrane to obtain the fermentation broth, and concentrated to a relative density of 1.05-1.1.

[0020] Furthermore, in the above-mentioned natural anti-hair loss composition, the fermentation process in step (c) is controlled in two stages:

[0021] Phase 1 (0-48 hours): maintain pH 5.5-6.0, dissolved oxygen ≤ 5%;

[0022] The second stage (48-96 hours): adjust the pH to 4.0-4.5 and the dissolved oxygen content ≤2%.

[0023] Furthermore, in the above-mentioned natural anti-hair loss composition, the weight ratio of the components in the natural carrier is:

[0024] Vegetable glycerin: cold-pressed camellia oil: algin = (6-8): (1-2): (0.1-0.3), where:

[0025] Vegetable glycerin purity ≥99%;

[0026] The 6-shogaol content of cold-pressed camellia oil is ≥ 0.5%;

[0027] The fucoidan content in alginate is ≥80%.

[0028] The present invention also discloses a method for preparing the composition, comprising the following steps:

[0029] (1) Mix the Chinese patent medicine active ingredients and the plant active ingredients at 35-45°C;

[0030] (2) adding the polypeptide complex and stirring at a low speed for 20-30 min, wherein the stirring speed is ≤200 rpm;

[0031] (3) Preheating the natural carrier to 40-50° C. and then mixing it into (2) and stirring at 1500-2000 rpm×5-10 min to obtain the composition.

[0032] Furthermore, in the preparation method of the above composition, natural vitamin E is added as an antioxidant in step (3) at a final concentration of 0.1-0.3 wt%.

[0033] In a second aspect, the present invention also discloses the use of the above composition in the preparation of anti-hair loss products.

[0034] In a third aspect, the present invention further discloses the use of the above composition in combination with minoxidil in the preparation of an anti-hair loss product.

[0035] In a fourth aspect, the present invention further discloses an anti-hair loss product comprising any one of the above compositions and minoxidil at a final concentration of 0.25-1.0 wt%.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] The present invention discloses a composition containing polypeptides. The composition has high safety, abandons chemical excipients such as propylene glycol and parabens, and adopts natural carriers such as vegetable glycerin and camellia oil. Human trials show that the adverse reaction rate is only 2.5%, which is much lower than that of commercially available minoxidil products. At the same time, the composition also has good antibacterial properties. GHK-Cu and palmitoyl tripeptide-1 promote the proliferation of hair papilla cells and upregulate VEGF expression, while acetyl tetrapeptide-3 inhibits 5α-reductase activity and reduces DHT damage to hair follicles. When used in combination with active ingredients of Chinese herbal medicines, the hair regeneration rate is increased to 89.3%.

[0038] The present invention adopts two-stage fermentation and vacuum low-temperature concentration, with a total energy consumption of 3.2kW·h / kg, which is only 36% of the traditional high-temperature extraction process (8.9kW·h / kg). It is green and sustainable. In addition, the water extraction and alcohol precipitation process is used throughout the process, with an active ingredient retention rate of >85%, an ethanol recovery rate of ≥85%, and VOCs emissions of only 48ppm.

[0039] The present invention enhances the penetration of fat-soluble components by using camellia oil (oleic acid ≥ 80%), and alginate forms a sustained-release gel layer, so that the cumulative permeation amount of ginsenoside Rg1 in 24 hours reaches 38.2 μg / cm 2 .

[0040] When the composition of the present invention is used in combination with 0.5% minoxidil, the hair regeneration rate reaches 93.6% without irritation. In addition, minoxidil is pre-dissolved in camellia oil to avoid complex precipitation with the polypeptide, thereby ensuring the uniformity of the preparation. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] In order to more clearly illustrate the technical solution of the present invention, the following is a brief introduction to the drawings required for use in the implementation. Obviously, the drawings described below are only some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0042] Figure 1 The results of the transdermal absorption rate test of ginsenoside Rg1 in Test Example 1 are shown.

[0043] Figure 2 The GHK-Cu transdermal absorption rate test results in Test Example 1.

[0044] Figure 3 The hair regrowth rate test results in Test Example 2.

[0045] Figure 4 The hair follicle density test results in Test Example 2. DETAILED DESCRIPTION

[0046] The following is a clear and complete description of the technical solutions in the embodiments of the present invention in conjunction with specific embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0047] In the present invention, when referring to numerical ranges, unless otherwise specified, the numerical ranges are considered continuous and include the minimum and maximum values of the range, as well as every value between such minimum and maximum values. Furthermore, when a range refers to an integer, every integer between the minimum and maximum values of the range is included. In addition, when multiple ranges are provided to describe a feature or characteristic, the ranges can be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges subsumed therein.

[0048] In the present invention, the raw materials and instruments involved are shown in the following table:

[0049] Table 1 Source of raw materials

[0050]

[0051]

[0052] Table 2 Instruments and Equipment

[0053]

[0054]

[0055] Example 1

[0056] A composition containing a polypeptide, comprising:

[0057] 1. Components:

[0058] Active ingredients of Chinese patent medicine: 70 parts (Polygonum multiflorum: Platycladus orientalis leaves: Ligustrum lucidum fruit: Eclipta prostrata = 10:15:8:7)

[0059] Plant active ingredients: 30 parts (ginseng fermentation liquid: Ganoderma lucidum polysaccharide = 2.5:1)

[0060] Peptide complex: 5 parts (GHK-Cu: Palmitoyl Tripeptide-1: Acetyl Tetrapeptide-3 = 1:1:0.5) Natural carrier: 100 parts (Vegetable Glycerin: Cold-pressed Camellia Oil: Alginate = 7:1.5:0.2)

[0061] 2. Preparation method:

[0062] 1) Preparation of active ingredients of Chinese patent medicines:

[0063] Weigh 10 kg of Polygonum multiflorum, 15 kg of Platycladus orientalis leaves, 8 kg of Ligustrum lucidum fruits, and 7 kg of Eclipta prostrata, and crush them into 20-mesh particles.

[0064] Add 10 times the amount (400 kg) of purified water, boil in a stainless steel extraction tank (100 ± 2 ° C), and maintain a slight boiling temperature for 3 times, each time for 1.5 hours.

[0065] The three extracts were combined, filtered through a 200-mesh filter cloth, and the filtrate was transferred to a vacuum concentrator (vacuum degree -0.08 MPa, temperature 60°C) and concentrated to a relative density of 1.18 (measured at 60°C).

[0066] 95% ethanol was added to a final concentration of 63% (ethanol volume fraction), stirred for 30 minutes, and allowed to stand for 24 hours.

[0067] The supernatant was collected and centrifuged at 5000 g for 15 minutes (centrifuge model: H-2050R, Hunan Xiangyi), and concentrated until there was no alcohol taste, to obtain the active ingredient extract of the Chinese patent medicine (solid content 65%).

[0068] 2) Preparation of ginseng fermentation liquid:

[0069] Fresh ginseng was washed and sliced (thickness 2-3 mm), mixed with sterile water at a ratio of 1:4 (w / v), and sucrose was added to adjust the sugar content to 10°Bx (saccharimeter: PAL-1, Atago, Japan).

[0070] Lactobacillus plantarum (CICC 20265, China Industrial Microbiology Culture Collection Center) was inoculated to a final concentration of 1×10 6 CFU / mL.

[0071] Two-stage fermentation control:

[0072] Phase 1 (0-48 hours): 37°C fermentation tank ( B Plus, Sartorius, Germany) was placed in a nitrogen atmosphere to maintain dissolved oxygen ≤ 5%, and 1 M NaOH was automatically added to maintain pH 5.8.

[0073] The second stage (48-96 hours): the temperature was adjusted to 40° C., nitrogen was introduced until the dissolved oxygen content was ≤2%, and 1 M HCl was added to maintain the pH at 4.3.

[0074] The fermentation broth was centrifuged at 5000g for 20 min, and the supernatant was filtered through a 0.45 μm filter (Millipore) and concentrated to a density of 1.08 (rotary evaporator, 40° C.).

[0075] 3. Preparation of composition:

[0076] 1) 70 parts of Chinese patent medicine active ingredient extract and 30 parts of plant active ingredients (21.4 parts of ginseng fermentation liquid + 8.6 parts of Ganoderma lucidum polysaccharide) were mixed in a 40°C water bath and magnetically stirred (600 rpm, 30 minutes).

[0077] 2) Add 5 parts of a polypeptide complex (2.0 parts of GHK-Cu, 12.0 parts of palmitoyl tripeptide, and 31.0 parts of acetyl tetrapeptide) and mix at low speed using a paddle stirrer (200 rpm, 30 minutes).

[0078] 3) Preheat the natural carrier (70 parts of vegetable glycerin, 15 parts of camellia oil, and 2 parts of alginate) to 45° C. and slowly add the above mixture. Homogenize at high speed (2000 rpm, 10 minutes, homogenizer T25, IKA, Germany).

[0079] 4) Add 0.2% natural vitamin E (Sigma-Aldrich), filter through a 0.22 μm sterile filter membrane, and dispense into light-proof bottles.

[0080] Examples 2-5

[0081] The component adjustments of other examples are detailed in Table 3 below. The preparation steps are the same as in Example 1, with only the parameters changed.

[0082] Table 3 Parameter setting table for Examples 2-5

[0083]

[0084] Example 4 Fermentation control: stage 1 (0-24 h, pH 6.0, dissolved oxygen ≤ 5%); stage 2 (24-72 h, pH 4.5, dissolved oxygen ≤ 2%); stage 3 (72-96 h, pH 4.0, dissolved oxygen ≤ 1%).

[0085] Example 5: Preparation of Minoxidil

[0086] In step 3 of Example 1, minoxidil (Sigma, purity ≥99%) was pre-dissolved in camellia oil (40° C.) and added simultaneously with the natural carrier to a final concentration of 0.5 wt %. The subsequent steps were the same as in Example 1.

[0087] Comparative Examples 1-6

[0088] The component adjustments of Comparative Examples 1-6 are detailed in Table 4 below.

[0089] Table 4 Parameter setting table of comparative examples 1-6

[0090]

[0091] Test Example 1

[0092] The transdermal absorption rates of different compositions were analyzed.

[0093] Samples: Examples 1-5, Comparative Examples 1-6, and a control group (commercially available minoxidil tincture).

[0094] In vitro pig skin treatment: fresh pig ear skin (thickness 0.8±0.1 mm) was taken, hair removed and then immersed in PBS (pH 7.4) for 2 hours.

[0095] Franz diffusion cell: effective diffusion area 2.54cm 2 The receiving solution was PBS containing 0.5% SDS (37°C constant temperature water bath).

[0096] Administration: Take 0.2 mL of sample and apply it evenly on the skin surface. Take 1 mL of sample at 0, 2, 4, 8, 12, and 24 hours (make up an equal amount of fresh solution).

[0097] Detection method:

[0098] Ginsenoside Rg1: HPLC (Agilent 1260, C18 column, mobile phase acetonitrile-water gradient elution, detection wavelength 203 nm).

[0099] GHK-Cu: ICP-MS (Agilent 7900, determination of Cu 2+ concentration, converted to peptide content).

[0100] Statistics:

[0101] Cumulative permeation (Q n )=(C n ×V+ΣC (n-1) ×V s ) / A, where V = volume of receiving cell (7 mL), V s = sampling volume (1 mL), A = diffusion area.

[0102] The results are expressed as mean ± SD (n = 6). One-way ANOVA was used for comparison among the groups, and p < 0.05 was considered a significant difference.

[0103] Table 5 Transdermal absorption rate test results

[0104]

[0105]

[0106] From Table 5 and Figure 1 、 Figure 2 It can be seen that the permeation amount of ginsenoside Rg1 in Example 1 (38.2 μg / cm 2 ) was significantly higher than that of comparative example 1 (12.4 μg / cm 2 ), mainly because the unsaturated fatty acids (oleic acid ≥ 80%) in camellia oil can enhance the fluidity of the stratum corneum, while the alginate forms a hydrated gel layer to delay the evaporation of active ingredients (p < 0.01, t test).

[0107] The GHK-Cu permeation rate of Example 5 (14.9 μg / cm 2 ) is slightly lower than Example 1 (15.6 μg / cm 2 ), which may be due to the slight competition of minoxidil for carrier binding sites, but the difference was not statistically significant (*p=0.12*).

[0108] Comparative Example 2 (no alcohol precipitation) had flavonoid impurities that blocked the pores, resulting in a GHK-Cu permeation rate of only 5.1 μg / cm 2 .

[0109] Test Example 2

[0110] The anti-hair loss efficacy of different compositions was analyzed.

[0111] Animals: C57BL / 6 mice (male, 8 weeks old), 10 mice per group, back hair removal (4 cm 2 ).

[0112] Dosing: Apply 0.1 mL of the sample daily for 21 days.

[0113] Detection indicators:

[0114] Hair regrowth rate: ImageJ was used to analyze the melanin coverage (%) of the depilated area.

[0115] Hair follicle counting: HE-stained sections (3 fields of view per sample, 200× microscopic imaging).

[0116] Table 6 Anti-hair loss efficacy test results

[0117]

[0118]

[0119] Note: “**” indicates p < 0.01 compared with the control group.

[0120] From Table 6 and Figure 3 、 Figure 4 It can be seen that the hair follicle density of Example 1 (32.1 / mm 2 ) Comparative Example 3 (peptide imbalance group, 21.5 / mm 2 ) was 49.3% higher, due to GHK-Cu promoting dermal papilla cell proliferation and palmitoyl tripeptide-1 upregulating VEGF expression (Western blot verification). Comparing Example 1 with Comparative Example 6 (unfermented ginseng), the hair regrowth rate increased by 53.3%, due to fermentation converting ginsenoside Rb1 into the more easily absorbed Rg3.

[0121] The hair regrowth rate of Example 5 (93.6%) exceeded that of the minoxidil alone group (76.4%) because minoxidil dilates blood vessels and the composition of the present invention prolongs the growth phase of hair follicles (*enhanced staining of hair follicle cycle marker Ki-67*).

[0122] Test Example 3

[0123] Analyze the long-term effectiveness and safety of different compositions.

[0124] Participants: 120 patients with androgenic alopecia (grade II-IV) were randomly divided into 6 groups for double-blind control.

[0125] Protocol: Apply 1 mL of sample every morning and evening for 6 months.

[0126] Evaluation Metrics:

[0127] Adverse reaction rate: erythema, itching, and desquamation were recorded.

[0128] Hair density: Image analysis (roots / cm 2 ).

[0129] Table 7 Long-term effectiveness and safety test results

[0130]

[0131]

[0132] As shown in Table 7, the adverse reaction rate of Comparative Example 1 (containing propylene glycol) was 28.6%, mainly because propylene glycol damaged the skin barrier (transepidermal water loss value TEWL increased by 35%). <51> ) still meets the standard, this is because the 6-gingerol in camellia oil has natural antibacterial properties (inhibition zone diameter ≥ 8mm).

[0133] The hair density improvement of Example 5 (42.1%) was significantly higher than that of commercially available minoxidil (31.5%), and there were no side effects such as scalp itching (*patient satisfaction score 4.7 / 5 vs 3.2 / 5*).

[0134] Test Example 4

[0135] The different compositions were analyzed in an accelerated stability test.

[0136] Conditions: 40℃ / 75%RH, 6 months, monthly sampling and testing.

[0137] Detection method:

[0138] Flavonoid retention rate: UV method (rutin standard curve, 510 nm).

[0139] Polysaccharide retention rate: phenol-sulfuric acid method (490 nm).

[0140] Table 8 Stability accelerated test results

[0141]

[0142]

[0143] Note: The control group was a commercially available plant anti-hair loss essence.

[0144] As shown in Table 8, the flavonoid retention rate of Example 1 (91.3%) is much higher than that of Comparative Example 2 (63.4%), because the alcohol precipitation removes polysaccharide impurities and reduces the degradation caused by the Maillard reaction.

[0145] The polysaccharide retention rate of Comparative Example 4 (addition of methylparaben) (76.9%) was lower than that of Example 1 (88.7%) because the chemical preservative accelerated the breakage of the alginate chains (the molecular weight measured by GPC decreased by 23%).

[0146] The flavonoid retention rate of the commercially available control group (high-temperature extraction process) was only 72.5%, confirming the low-temperature advantage of the two-stage fermentation of the present invention (*calculated activation energy Ea=45kJ / mol vs. traditional process Ea=68kJ / mol*).

[0147] Test Example 5

[0148] The process environmental friendliness of the composition was analyzed.

[0149] VOCs detection: GC-MS (Agilent 7890B / 5977A) headspace sampling to determine residual ethanol, propylene glycol, etc.

[0150] Energy consumption calculation: total power consumption of extraction, concentration, and fermentation processes (kW·h / kg product).

[0151] Table 9 Process environmental protection test results

[0152] Group Total VOCs (ppm) Energy consumption (kW·h / kg) Example 1 48±5** 3.2±0.3** Comparative Example 1 520±30 4.8±0.5 Comparative Example 2 280±15 7.1±0.6 Commercially available process 650±50 8.9±0.7

[0153] Note: indicates p<0.01 compared with the commercial process.

[0154] As shown in Table 9, the total VOCs content of Example 1 (48 ppm) is only 7.4% of that of the commercial process (650 ppm) because water extraction and alcohol precipitation are used throughout the process, avoiding organic solvents such as methanol (*GC-MS detection of ethanol residue <0.1%*).

[0155] The energy consumption of Example 1 (3.2 kW·h / kg) is 64% lower than that of the conventional process (8.9 kW·h / kg), mainly because vacuum concentration (60°C) replaces atmospheric pressure concentration (90°C), and the energy saving rate reaches 42%.

[0156] The COD value of the process wastewater of the present invention (320 mg / L) is much lower than that of comparative example 2 (1200 mg / L) because the ethanol precipitated from the alcohol can be recovered and reused (recovery rate ≥85%).

[0157] In summary, the present invention provides a composition containing polypeptides through precise component combination (synergistic combination of Chinese herbal medicine, polypeptides, and fermentation active ingredients) and green process innovation (two-stage fermentation + alcohol precipitation purification). Its transdermal efficacy is enhanced by 3.1 times, and the synergistic effect of polypeptides increases the hair follicle density by 32.1 / mm 2 The 6-month adverse reaction rate is only 2.5%, the flavonoid retention rate is >90%, VOCs emissions are reduced by 90%, and the energy consumption is only 36% of the traditional process. It avoids the problems of existing technologies inhibiting hair follicle activation signal pathways, insufficient hair follicle density improvement, and decreased flavonoid retention rate. It achieves a triple breakthrough in efficacy, safety, and sustainability, which is conducive to the development of anti-hair loss technology and provides new ideas for anti-hair loss products.

[0158] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

Claims

1. A composition containing a polypeptide, characterized in that It is composed of the following components in parts by weight: Active ingredient of Chinese patent medicine: 40-100 parts, prepared by mixing Polygonum multiflorum, Platycladus orientalis leaves, Ligustrum lucidum fruits, and Eclipta prostrata in a weight ratio of (5-15):(8-20):(3-12):(4-10), followed by water extraction and alcohol precipitation; Plant active ingredients: 15-50 parts, including ginseng fermentation liquid and Ganoderma lucidum polysaccharide; Peptide complex: 1-10 parts, composed of copper peptide GHK-Cu, palmitoyl tripeptide-1, and acetyl tetrapeptide-3 in a weight ratio of (1:0.5-2:0.3-1); Natural carrier: 50-150 parts, including vegetable glycerin, camellia oil and algae gum.

2. The composition according to claim 1, characterized in that The preparation method of the active ingredient of the Chinese patent medicine is as follows: The Chinese patent medicine forms of Polygonum multiflorum, Platycladus orientalis leaves, Ligustrum lucidum fruits and Eclipta prostrata are mixed in proportion, 5-15 times the weight of water are added and boiled for extraction 2-4 times, each time for 1-2 hours, and the combined extracts are concentrated to a relative density of 1.15-1.20, which is measured at 60°C, and precipitated with alcohol to a final concentration of 60-65% ethanol.

3. The composition according to claim 1, characterized in that Among the plant active ingredients: The weight ratio of ginseng fermentation liquid to ganoderma lucidum polysaccharide is (2-3):1, and the preparation method of the ginseng fermentation liquid comprises the following steps: (a) Wash and slice fresh ginseng, then mix with sterile water at a ratio of 1:3-5 (w / v); (b) adding 3-5% sucrose solution and adjusting the sugar content of the final solution to 8-12°Bx; (c) Inoculate Lactobacillus plantarum to a final concentration of 1×10 6 CFU / mL, fermented at 37-40°C for 72-96 hours; (d) The supernatant was collected by centrifugation at 3000-5000 g, filtered through a 0.45 μm membrane to obtain the fermentation broth, and concentrated to a relative density of 1.05-1.

1.

4. The composition according to claim 3, characterized in that The fermentation process in step (c) is controlled in two stages: Phase 1 (0-48 hours): maintain pH 5.5-6.0, dissolved oxygen ≤ 5%; The second stage (48-96 hours): adjust the pH to 4.0-4.5 and the dissolved oxygen content ≤2%.

5. The composition according to claim 1, characterized in that The weight ratio of each component in the natural carrier is: Vegetable glycerin: cold-pressed camellia oil: algin = (6-8): (1-2): (0.1-0.3), where: Vegetable glycerin purity ≥99%; The 6-shogaol content of cold-pressed camellia oil is ≥ 0.5%; The fucoidan content in alginate is ≥80%.

6. The method for preparing the composition according to any one of claims 1 to 5, wherein: The following steps are involved: (1) Mix the Chinese patent medicine active ingredients and the plant active ingredients at 35-45°C; (2) adding the polypeptide complex and stirring at a low speed for 20-30 min, wherein the stirring speed is ≤200 rpm; (3) Preheating the natural carrier to 40-50° C. and then mixing it into (2) and stirring at 1500-2000 rpm×5-10 min to obtain the composition.

7. The method according to claim 6, characterized in that In step (3), natural vitamin E is added as an antioxidant at a final concentration of 0.1-0.3 wt%.

8. Use of the composition according to any one of claims 1 to 5 in the preparation of an anti-hair loss product.

9. Use of the composition according to any one of claims 1 to 5 in combination with minoxidil in the preparation of an anti-hair loss product.

10. An anti-hair loss product, characterized in that: Contains the composition according to any one of claims 1 to 5 and minoxidil with a final concentration of 0.25-1.0 wt%.

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