Amino acid shampoo with hair loss preventing, moisturizing and repairing effects and preparation method of amino acid shampoo

Abstract

The invention discloses amino acid shampoo and a preparation method thereof. The shampoo comprises an amino acid surfactant, polypeptide A, a composite plant extract, a fermentation extract, hyaluronic acid modified molecules and the like. Through scientific and optimized proportioning and preparation process, the shampoo has mild property, high cleaning power and moisturizing and repairing effects. The polypeptide A promotes proliferation of hair follicle dermal papilla cells by regulating a Wnt / beta-catenin signal channel, and the composite plant extract and the fermentation extract have a synergistic effect, so that the scalp repair and anti-inflammatory effects are further enhanced. The shampoo is excellent in biodegradability, mild and low in irritation, and is suitable for improving scalp health and promoting hair regeneration.

Description

Technical Field

[0001] The invention relates to the technical field of daily chemical products, and in particular to an amino acid shampoo with hair loss prevention, moisturizing and repairing effects and a preparation method thereof. Background Art

[0002] The existing anti-hair loss shampoos on the market mainly work by adding chemical synthetic ingredients (such as selenium sulfide, imidazole drugs, etc.). Although these ingredients can alleviate the problem of hair loss in the short term, they may cause side effects such as scalp irritation and allergies. At the same time, long-term use may also lead to dependence and hair loss rebound problems. In addition, although ingredients such as silicone oil added to some shampoos can temporarily improve the smoothness of hair, long-term use can easily lead to clogging of scalp pores, thereby exacerbating the problem of hair loss.

[0003] On the other hand, dry, frizzy and split hair are also common hair problems that consumers are concerned about. The moisturizing ingredients in traditional shampoos are mainly glycerin and propylene glycol, but the moisturizing effect of these ingredients is relatively simple and cannot meet the needs of deep hair repair caused by environmental factors or dyeing and perming damage. In addition, the detergents contained in most shampoo products are traditional sulfate surfactants (such as sodium lauryl sulfate, SLS). Although they have strong cleaning ability, they are very irritating to the scalp and hair, and can easily damage the natural barrier of the scalp, thereby causing dryness, sensitivity and even inflammation of the scalp, further aggravating hair damage.

[0004] In recent years, amino acid surfactants have gradually become a popular choice for shampoo products due to their mildness and protective effect on the skin barrier. However, existing amino acid shampoos are mainly used for basic cleaning and improving sensitive scalps, lacking targeted functional designs. In the field of functional shampoos, how to effectively combine anti-hair loss with deep moisturizing and repairing functions, and achieve a balance between the mildness and safety of the formula, is still a technical problem that needs to be solved urgently. Based on this, the present invention combines active polypeptides, plant extracts and high-efficiency moisturizing factors to develop an amino acid shampoo with both anti-hair loss and moisturizing and repairing functions to meet the multiple needs of consumers while avoiding the deficiencies in the prior art. Summary of the invention

[0005] The present invention aims to provide a mild, low-irritation, high-cleansing amino acid shampoo, and at the same time enhance the hair follicle repair and scalp health improvement effects of the shampoo by adding functional active ingredients such as polypeptide A, compound plant extracts and fermented extracts.

[0006] Therefore, the present invention discloses an amino acid shampoo on one hand, wherein the shampoo comprises the following components per 1000 g:

[0007] (1) an amino acid surfactant, wherein the amino acid surfactant is an AAS1 complex: 8%-12%;

[0008] (2) Polypeptide A: 0.1%-0.3%;

[0009] (3) Complex plant extracts: 2%-4%;

[0010] (4) Fermentation extract: 1%-2%;

[0011] (5) Hyaluronic acid modified molecules: 0.3%-0.5%;

[0012] (6) a thickener, wherein the thickener is xanthan gum: 0.5%-1.5%;

[0013] (7) pH adjuster and deionized water: The pH adjuster is lactic acid with a pH value of 5.0-5.5, and the balance is made up of deionized water to 100%.

[0014] In one aspect, the present invention further discloses an amino acid shampoo, wherein 1000 g of the shampoo comprises the following components:

[0015] (1) amino acid surfactant, wherein the amino acid surfactant is AAS1 complex: 100 g;

[0016] (2) Polypeptide A: 2 g;

[0017] (3) Complex plant extract: 30 g;

[0018] (4) Fermentation extract: 15 g;

[0019] (5) Hyaluronic acid modified molecules: 4 g;

[0020] (6) thickener, wherein the thickener is xanthan gum: 10 g;

[0021] (7) pH adjuster and deionized water: Use appropriate amount of lactic acid to adjust the pH to 5.0-5.5, and make up to 1000 g with deionized water.

[0022] Preferably, the AAS1 complex of the present invention comprises the following components:

[0023] (1) Sodium cocoyl glutamate: 500g;

[0024] (2) Potassium cocoyl glycinate: 300 g;

[0025] (3) Sodium cocoyl alanine: 100g;

[0026] (4) Sodium cocoyl isethionate: 50 g;

[0027] (5) Glycerol: 30 g;

[0028] (6) Citric acid: Use appropriate amount of citric acid to adjust pH to 5.0-6.0;

[0029] (7) Deionized water: Make up to 1000 g with deionized water.

[0030] Preferably, the amino acid sequence of polypeptide A of the present invention is KGPVQCGFSK, and its molecular weight is about 1175.4 Da.

[0031] Preferably, the composite plant extract of the present invention comprises active ingredients obtained from Polygonum multiflorum and Camellia sinensis.

[0032] Preferably, the total sugar content of the fermentation extract of the present invention is 25.6±0.8 mg / mL; 90% of the fermentation extract is oligosaccharides, wherein the molecular weight of the oligosaccharides is <1 kDa, and the purity of the oligosaccharides is 98.3±0.4%; wherein the main components of the oligosaccharides are L-fucose, D-galactose, and mannose, and the molar ratio of L-fucose, D-galactose, and mannose is 3:2:1.

[0033] Preferably, the molecular weight of the hyaluronic acid modified molecules of the present invention is ≤0.5 kDa on average, with a distribution range of 0.3-0.5 kDa, accounting for ≥90%.

[0034] In one aspect, the present invention also discloses a method for preparing the shampoo, the method comprising the following steps:

[0035] (1) Dissolving the basic cleaning system: dissolve the AAS1 complex in 50°C deionized water and stir for 30 minutes to form a basic cleaning solution A; (2) Active substance integration: dissolve polypeptide A, compound plant extract and fermented extract separately, add them to solution A one by one, and keep stirring at a low speed; (3) Adding the moisturizing and repairing system: dissolve the hyaluronic acid modified molecules in a small amount of water, slowly add them to the system, and control the temperature below 35°C; (4) Thickening and adjustment: add xanthan gum and adjust the pH to 5.0-5.5 with lactic acid, then use deionized water to make up the volume, stir evenly, filter and fill; (5) Finished product packaging: filter the final solution to ensure that there are no bubbles, fill it into a sterile container and seal it for storage.

[0036] In one aspect, the present invention also discloses a use of the AAS1 complex, polypeptide A, composite plant extract, fermentation extract and hyaluronic acid modified molecule in the preparation of amino acid shampoo.

[0037] Compared with the prior art, the amino acid shampoo of the present invention has the following advantages: (1) Gentle cleansing: It adopts an amino acid-derived surfactant complex system to reduce scalp irritation; (2) Promotes hair follicle regeneration: Polypeptide A significantly promotes the proliferation of hair follicle dermal papilla cells through the Wnt / β-catenin signaling pathway; (3) Improves the scalp environment: The composite plant extract and fermentation extract synergistically repair the scalp barrier, and have anti-inflammatory and antioxidant effects; (4) Ecological and environmental protection: It has excellent biodegradability and meets environmental protection requirements. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 Comparison results before and after use of the shampoo of the present invention. DETAILED DESCRIPTION

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0040] Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art. Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0041] Example 1: Formula and preparation of amino acid shampoo

[0042] 1. The raw material ratio of amino acid shampoo is as follows (taking 1000g shampoo as an example):

[0043] (1) Amino acid surfactant (AAS1 complex, see Example 2 for preparation): 8%-12%;

[0044] (2) Active ingredients:

[0045] 1) Polypeptide A (see Example 3 for details): 0.1%-0.3%,

[0046] 2) Composite plant extract (see Example 4 for details): 2%-4%,

[0047] 3) Fermentation extract (see Example 5 for details): 1%-2%;

[0048] (3) Hyaluronic acid modified molecules (see Example 6 for details): 0.3%-0.5%;

[0049] (4) Auxiliary ingredients:

[0050] 1) pH adjuster (lactic acid): appropriate amount,

[0051] 2) Thickener (xanthan gum): 0.5%-1.5%,

[0052] 3) Deionized water: make up to 100%.

[0053] 2. According to the above formula, the raw material ratio of the amino acid shampoo used in the present invention is as shown in Table 1:

[0054] Table 1 Raw material ratio of amino acid shampoo

[0055]

[0056] 3. Preparation of amino acid shampoo

[0057] (1) Dissolving the basic cleaning system: dissolving the AAS1 complex in 50°C deionized water and stirring for 30 minutes to form a basic cleaning solution A;

[0058] (2) Active substance integration: Dissolve polypeptide A, compound plant extract and fermentation extract separately and add them to solution A one by one while stirring at a low speed;

[0059] (3) Addition to the moisturizing and repairing system: Dissolve the hyaluronic acid modified molecules in a small amount of water and slowly add them to the system, keeping the temperature below 35°C;

[0060] (4) Thickening and adjustment: add xanthan gum and adjust the pH to 5.0-5.5 with lactic acid, then use deionized water to make up the volume, stir evenly, filter and fill;

[0061] (5) Finished product packaging: The final solution is filtered to ensure there are no bubbles, and then filled into a sterile container and sealed for storage.

[0062] Example 2: Preparation and testing of AAS1 complex

[0063] 1. Formula of AAS1 complex: A compound system based on amino acid-derived surfactant is prepared to meet the requirements of mildness, low irritation and high cleaning power. The specific formula is shown in Table 2:

[0064] Table 2 Formulation of AAS1 complex

[0065] Element Content (g) Function Sodium Cocoyl Glutamate 500 Gentle cleansing and moisturizing Potassium Cocoyl Glycinate 300 Fine foam, low irritation Sodium Cocoyl Alanine 100 Cleansing enhancement, softening performance Sodium Cocoyl Isethionate 50 Stable foam, enhanced cleaning power glycerin 30 Moisturizer Citric acid (to adjust pH) Moderate Maintain the solution acid-base balance, pH 5.0-6.0 Deionized water Top up to 1000 Solvents

[0066] 2. Preparation process of AAS1 complex

[0067] (1) Prepare the base solution: Add deionized water (60°C) to a stirring kettle and keep stirring at a low speed. Gradually add sodium cocoyl glutamate and stir until it is completely dissolved to form a transparent and uniform solution A.

[0068] (2) Dissolving auxiliary surfactants: Dissolve potassium cocoyl glycinate and sodium cocoyl isethionate in warm water (50°C) in proportion and stir evenly to form solution B.

[0069] (3) Combine the solutions: Slowly pour solution B into solution A, stirring continuously to avoid lumps or stratification. Add sodium cocoyl alanine and continue stirring until the solution is completely homogenized.

[0070] (4) Adjust the system: Cool down to 40°C, slowly add glycerin, and stir for 10 minutes to enhance the moisturizing performance. Use citric acid to adjust the pH of the solution to between 5.0 and 6.0, and stir evenly.

[0071] (5) Filtration and packaging: Pass the solution through a 200-mesh filter to remove insoluble impurities. Place the solution in a sealed container and store at room temperature for later use.

[0072] 3. Comparative examination of AAS1 complex

[0073] Product A: Sodium cocoyl glutamate + potassium cocoyl glycinate; manufactured by Ajinomoto.

[0074] Product B: Sodium cocoyl sarcosinate + sodium cocoyl isethionate; manufactured by Clariant.

[0075] Product C: Sodium cocoyl alanine single ingredient; manufactured by Stepan.

[0076] (1) Appearance and stability: The appearance changes of the samples were observed at 40°C and in a frozen environment. The results (Table 3) showed that the stability of the AAS1 complex in high temperature and frozen environments was better than that of product B, and was comparable to that of products A and C.

[0077] Table 3 Comparison results of appearance and stability

[0078] project AAS1 complex Product A Product B Product C Room temperature appearance Transparent liquid Transparent liquid Light yellow liquid Transparent liquid Stability at 40℃ No stratification, no precipitation No stratification, no precipitation Slight precipitation No stratification, no precipitation Freeze stability No turbidity No turbidity Crystallization No turbidity

[0079] (2) pH value: The pH value of the 1% solution was measured. The results (Table 4) showed that the AAS1 complex was consistent with commercial products A and B, and met the weakly acidic environment requirements of human skin.

[0080] Table 4 pH value test results

[0081] project AAS1 complex Product A Product B Product C pH Range 5.0-5.5 5.0 5.2 5.8

[0082] (3) Surface activity (CMC test): The CMC value (critical micelle concentration) was measured by a surface tension meter. The results showed (Table 5) that the AAS1 complex had the lowest CMC value and the strongest surface activity, which was superior to all control products.

[0083] Table 5 Surface activity test results

[0084] project AAS1 complex Product A Product B Product C CMC value (%) 0.03 0.05 0.07 0.05 Surface tension (mN / m) 26.5 28.0 30.2 28.0

[0085] (4) Mildness test (cell survival rate): HaCaT cells were treated with 1% solution and the cell survival rate was detected. The results showed (Table 6) that the AAS1 complex had the highest mildness and had no obvious toxicity to cells.

[0086] Table 6 Mildness test results

[0087] project AAS1 complex Product A Product B Product C Cell survival rate (%) 95.6 93.2 91.5 92.8

[0088] (5) Foaming performance: The foaming volume and stable foam volume of 1% solution were tested. The results showed (Table 7) that the foaming performance of the AAS1 complex was significantly better than that of products A and B, and slightly higher than that of product C.

[0089] Table 7 Foaming performance test results

[0090] project AAS1 complex Product A Product B Product C Foaming volume (mL) 380 340 330 370 Stable bubble volume (mL) 360 320 300 350

[0091] (6) Biodegradability: The degradation rate within 28 days was tested according to the OECD 301B standard. The results show (Table 8) that the degradation rate of the AAS1 complex is the highest, which meets the environmental protection requirements.

[0092] Table 8 Biodegradability test results

[0093] project AAS1 complex Product A Product B Product C Degradation rate (%) 96.2 94.5 90.1 92.8

[0094] Example 3: Design and preparation of polypeptide A

[0095] 1. Design of polypeptide A: Polypeptide A (amino acid sequence is KGPVQCGFSK, molecular weight is about 1175.4Da) is designed based on the structure that promotes the proliferation of hair follicle dermal papilla cells (DPCs), aiming to promote hair follicle regeneration by regulating the Wnt / β-catenin signaling pathway. In this sequence, lysine (K) and glutamine (Q) enhance the affinity of the molecule for binding to LRP5 / 6, cysteine ​​(C) may provide stability of the molecular structure, and phenylalanine (F) and serine (S) promote downstream signal activation.

[0096] 2. Preparation method of polypeptide A: Use solid phase synthesis (SPPS) to gradually couple each amino acid monomer, and Wang resin is selected as the resin carrier. After the synthesis is completed, trifluoroacetic acid (TFA) is used to deprotect the group and release the polypeptide. Purify by high performance liquid chromatography (HPLC), the purity of the polypeptide is >98%. Freeze-dry and store for future use.

[0097] 3. Comparison of the biological effect experiment of polypeptide A with the control of commonly used polypeptides

[0098] The experimental groups are as follows: Experimental group 1: polypeptide A (100 μM); Control group 1: commercial polypeptide B (Peptide International Inc.) (sequence: Ac-EPRGGFGRC-NH2); Control group 2: commercial polypeptide C (Sigma-Aldrich) (sequence: Ac-KGHK-NH2); Blank group: no polypeptide treatment.

[0099] (1) DP cell proliferation assay

[0100] DP cells were seeded in 96-well plates at 5000 cells / well and replaced with serum-free medium after 24 hours of culture. Peptide samples were added: A, B, C, and the concentrations were set to 0μM (control), 1μM, 5μM, and 10μM. After 72 hours of culture, MTT reagent was added and incubated for 4 hours. The OD570 value was measured after DMSO dissolved the crystals. The effects of different peptides on the proliferation of DP cells were compared, and the proliferation rate at a concentration of 10μM was used as a reference point.

[0101] The results showed that the cell proliferation rate of polypeptide A at a concentration of 10 μM was 145%, which was significantly higher than that of polypeptide B (120%) and polypeptide C (110%) (P<0.05).

[0102] (2) Analysis of hair follicle-related gene expression

[0103] After DP cells were cultured to 70% confluence, 10 μM peptide samples were added for 24 hours. Total RNA was extracted and reverse transcribed into cDNA, and RT-qPCR was used to detect the expression levels of genes related to the Wnt signaling pathway (AXIN2, β-catenin). GAPDH was used as an internal reference gene to calculate the relative expression (2^-ΔΔCt). The upregulation effect of peptide A on gene expression was compared with that of peptides B and C to evaluate their ability to activate the Wnt / β-catenin pathway.

[0104] The results showed that polypeptide A significantly upregulated the expression of AXIN2 and β-catenin genes (3.2-fold and 2.8-fold of the control, respectively), which was higher than polypeptide B (2.1-fold and 1.9-fold) and polypeptide C (1.8-fold and 1.5-fold).

[0105] (3) In vitro hair follicle growth experiment

[0106] Hair follicles were isolated from the scalp tissue of volunteers, randomly divided into groups, and cultured in hair follicle growth medium. 10μM peptide samples were added for treatment, 6 hair follicles were set in each group, and cultured continuously for 14 days, and the medium was changed every 3 days. The changes in hair follicle length were recorded, and the hair follicle growth rate was calculated (each hair follicle growth exceeding 0.3mm was considered positive). The effects of different peptides on the promotion of hair follicle growth were compared, with the hair follicle growth rate and average length change as indicators.

[0107] The results showed that the hair follicle growth rate of polypeptide A group was 83.3%, significantly higher than polypeptide B (66.7%) and polypeptide C (50%), and the average length increased by 0.56mm, higher than polypeptide B (0.42mm) and polypeptide C (0.32mm).

[0108] Example 4: Preparation of composite plant extracts

[0109] 1. Detailed preparation method of extract

[0110] (1) Preparation of plant root extract

[0111] Take the root of Polygonum multiflorum (He Shouwu), dry it in the sun, crush it, and pass it through an 80-mesh sieve for later use. Take 1000g of the crushed root material, add 70% ethanol aqueous solution (liquid-to-solid ratio 10:1mL:g), reflux extraction at 85°C for 2 hours, and repeat the extraction 3 times. After the extracts are combined, use a rotary evaporator (40°C, 60rpm) to remove ethanol and concentrate it to a paste with a relative density of 1.2. Dissolve the concentrated paste in deionized water, pass it through an HP-20 macroporous resin column, wash the impurities with deionized water, and then use 30% ethanol and 70% ethanol gradient elution in turn, and collect the 70% ethanol elution portion. The target eluate is vacuum freeze-dried to obtain a light yellow powder, which is named plant extract A.

[0112] The detection by high performance liquid chromatography (HPLC) showed that the content of stilbene glycoside was ≥ 2%, and the content of epicatechin was about 0.8%.

[0113] (2) Preparation of plant leaf extracts

[0114] Select the tender leaves of Camellia sinensis (Camellia tree), dry them at low temperature, crush them, and pass them through an 80-mesh sieve for later use. Take 1000g of the crushed leaf raw material, add 70% ethanol aqueous solution (liquid-to-solid ratio 10:1mL:g), reflux extract at 80°C for 1.5 hours, and repeat the extraction twice. After the extracts are combined, the ethanol is removed by rotary evaporation and concentrated to a paste with a relative density of 1.1. The concentrated paste is dissolved in deionized water, passed through an HPD-100 macroporous resin column, first washed with deionized water, then eluted with 50% ethanol, and the target portion is collected. The eluate is vacuum freeze-dried to obtain a light green powder, which is named B plant extract.

[0115] HPLC determination of EGCG content ≥ 3%, total polyphenol content of about 4.5%.

[0116] (3) Preparation of A+B composite extract

[0117] Mix plant extract A and plant extract B in a mass ratio of 1:1, mix thoroughly and evenly, and then vacuum pack and store for later use.

[0118] 2. Comparative test with commercially available products

[0119] Product A: Polygonum multiflorum root extract (Shaanxi Tianxingjian Bioengineering Co., Ltd.), standardised to ≥ 1.5% stilbene glycosides.

[0120] Product B: Camellia sinensis leaf extract (Hunan Green Rhythm Biotechnology Co., Ltd.), EGCG standardized ≥ 2%.

[0121] Product C: composite plant extract (Shaanxi Ruimei Shenghe Biotechnology Co., Ltd.), total content of diphenylethylene glycosides + EGCG standardized ≥ 3%.

[0122] Experimental groups: experimental group (A+B composite extract prepared as above), control group (commodity A, commodity B, commodity C), blank group (without any treatment).

[0123] (1) Antioxidant test

[0124] The antioxidant capacity of each group was tested using the DPPH free radical scavenging experiment. The lower the IC50 value, the stronger the antioxidant capacity. The results showed that the IC50 of the experimental group was 12.5 μg / mL, and the IC50 of the commodity A, commodity B, and commodity C groups were 19.2 μg / mL, 21.7 μg / mL, and 15.4 μg / mL, respectively. Therefore, the composite extract of the present invention is significantly superior to commodities A, B, and C in antioxidant capacity.

[0125] (2) Cell protection experiment

[0126] H2O2 was used to induce oxidative stress damage in HaCaT cells, and the cell survival rate after treatment with the extract was detected (MTT method). The results showed that the cell survival rate of the experimental group was 92.3%, the cell survival rates of the product A, product B, and product C groups were 85.6%, 83.8%, and 88.4%, respectively, and the cell survival rate of the blank group was 58.2%. Therefore, the protective effect of the A+B composite extract on oxidative damage was significantly higher than that of the control group.

[0127] (3) Hair growth stimulation experiment

[0128] The effect of the extract on the proliferation of hair follicle cells was detected by human hair follicle fibroblast model (CCK-8 method). The results showed that the cell proliferation rate of the experimental group was 156.2%, the cell proliferation rates of product A, product B, and product C groups were 125.6%, 118.4%, and 134.7%, respectively, and the cell proliferation rate of the blank group was 0%. Therefore, the composite extract of the present invention has a significant advantage in promoting the proliferation of hair follicle cells and shows a good potential hair growth effect.

[0129] Example 5: Preparation of fermentation extract

[0130] 1. Material Preparation

[0131] Seaweed polysaccharide (Sigma-Aldrich); lactic acid bacteria Lactobacillus fermentum, strain number LF-201 (purchased from ATCC, model ATCC 23271); pH 6.5 phosphate buffer (10 g / L).

[0132] 2. Fermentation process

[0133] The strain is cultured in an MRS liquid culture medium for 24 hours until the OD600 reaches 1.5-2.0, which is used as a seed solution; 10 g / L seaweed polysaccharide is dissolved in a phosphate buffer, sterilized (121° C., 15 min), and cooled to room temperature; the lactic acid bacteria seed solution is inoculated into the seaweed polysaccharide solution at a ratio of 5% (v / v), and fermented at 37° C. and 150 rpm for 48 hours; the fermentation liquid is sterilized through a 0.22 μm filter membrane, and high molecular impurities are removed through ultrafiltration (MWCO 1 kDa), and the target product is concentrated and freeze-dried to obtain oligosaccharide powder.

[0134] 3. Product Analysis

[0135] (1) The total sugar content of the fermentation extract was determined using the phenol-sulfuric acid method and was found to be 25.6±0.8 mg / mL.

[0136] (2) Analysis of the molecular weight distribution of oligosaccharides by high-performance liquid chromatography-gel permeation chromatography (HPLC-GPC) showed that 90% of the fermentation extract was oligosaccharides, of which the molecular weight of the oligosaccharides was <1 kDa and the purity of the oligosaccharides was 98.3±0.4%. The main components were L-fucose, D-galactose, and mannose, and the molar ratio of L-fucose, D-galactose, and mannose was 3:2:1.

[0137] 4. Comparative experiment with existing commonly used products

[0138] Product A: Extracted from seaweed, the main component is medium molecular weight seaweed polysaccharide (2-10kDa). Manufacturer: CPKelco BioTech.

[0139] Product B: Oligosaccharides (molecular weight < 3 kDa) prepared by enzymatic hydrolysis. Manufacturer: Qingdao Mingyue Biomedical Materials Co., Ltd.

[0140] (1) Cell proliferation promotion experiment

[0141] HFDPC and HaCaT cells were seeded in 96-well plates at 2 × 10 4Cells were cultured for 24 hours. Different treatment groups were added: the extract of the present invention, commercial product A, and commercial product B (all at a concentration of 20 μg / mL). After 72 hours of culture, the cell proliferation was detected using CCK-8 reagent. The OD value was measured at 450 nm.

[0142] The results are shown in Table 9. The extract of the present invention has better effects on promoting the proliferation of hair follicle dermal papilla cells and keratinocytes than those of product A and product B.

[0143] Table 9 Cell proliferation promotion experimental results

[0144] Group HFDPC proliferation rate (%) HaCaT proliferation rate (%) Blank control group 100±5 100±3 Extract of the present invention 165±5 150±4 Product A 130±7 120±3 Product B 115±5 105±6

[0145] (2) Moisturizing effect experiment

[0146] EpiSkin kit was used to simulate the skin barrier, and the extract of the present invention, product A and product B (all 25 μg / mL) were added. After 48 hours of culture, the water loss rate (TEWL) of the skin barrier was measured. The percentage of water loss rate reduction was used as the moisturizing effect index.

[0147] The results are shown in Table 10. The water loss rate (TEWL) of the extracts of the present invention is lower than that of commodity A and commodity B.

[0148] Table 10 Moisturizing effect experimental results

[0149] Group TEWL reduction rate (%) Blank control group 0 Extract of the present invention 42±3 Product A 30±3 Product B 24±4

[0150] (3) Anti-inflammatory activity experiment

[0151] HaCaT cells were stimulated with 1 μg / mL LPS for 24 hours to induce an inflammatory response. The extract of the present invention, product A and product B (all 20 μg / mL) were added. After 48 hours of culture, the levels of IL-6 and TNF-α in the culture medium were detected using an ELISA kit.

[0152] The results are shown in Table 11. The extract of the present invention significantly reduces the secretion of IL-6 and TNF-α, which is 20%-35% higher than that of product A and product B, respectively.

[0153] Table 11 Anti-inflammatory activity experimental results

[0154] Group IL-6 reduction rate (%) TNF-α reduction rate (%) Blank control group 0 0 Extract of the present invention 65±4 60±4 Product A 45±3 40±3 Product B 30±2 25±3

[0155] Example 6: Preparation of Hyaluronic Acid Modified Molecules

[0156] 1. Preparation method: Dissolve high molecular weight hyaluronic acid in deionized water to prepare a solution with a concentration of 2% w / v, and stir thoroughly until completely dissolved. Add hyaluronidase, set the mass ratio of enzyme to substrate to 1:200, and mix well. Place the reaction mixture in a 37°C constant temperature water bath for 6 hours, stirring slightly every hour. The reaction solution is filtered through an ultrafiltration membrane (0.5kDa molecular weight cutoff) to remove macromolecular residues and collect the filtrate. The filtrate is analyzed by gel chromatography (GPC) to ensure that the molecular weight is concentrated in the range of 0.3-0.5kDa, accounting for ≥90%. The filtrate is freeze-dried to obtain LMW-HA powder, which is stored in a light-proof and dry environment for use.

[0157] 2. Through testing, the molecular weight of the modified hyaluronic acid molecule is ≤0.5kDa on average, with a distribution range of 0.3-0.5kDa, accounting for ≥90%. Compared with the original HMW-HA, the modified hyaluronic acid molecule has stronger scalp permeability and hair follicle cell affinity.

[0158] 3. Permeability experiment: A Franz diffusion cell was used to simulate the scalp permeation experiment. The hyaluronic acid modified molecules prepared by the present invention, product A (low molecular weight hyaluronic acid (molecular weight 0.5-1 kDa), Shandong Foster Biotechnology Co., Ltd.) and product B (high molecular weight hyaluronic acid (molecular weight>1.2 MDa), Huaxi Biotechnology Co., Ltd.) (each 2% w / v) were applied to the surface of the scalp barrier model, and the hyaluronic acid concentration in the diffusion liquid was detected after 12 hours.

[0159] The results showed that the osmotic concentrations of the hyaluronic acid modified molecules prepared by the present invention, product A and product B were 180±10 μg / cm 2 , 120±8μg / cm 2 、30±5μg / cm 2 This shows that the hyaluronic acid modified molecules prepared by the present invention show significant advantages in the scalp penetration experiment, which is 50% higher than that of product F and 6 times higher than that of product G.

[0160] Example 7: Application of amino acid shampoo

[0161] 1. Skin model verification of moisturizing and repairing effects

[0162] (1) Skin model preparation: A 3D human epidermal tissue model (EpiDerm) was used and the barrier function was destroyed by UV-B radiation.

[0163] (2) Experimental groups

[0164] Control group: no treatment;

[0165] Test group: the model was smeared with a dilution (1:10) of the shampoo of the present invention (prepared according to the formula of Section 2 of Example 1);

[0166] Existing product control group: The model was smeared with a diluted solution of commercial shampoo (Amino Mason amino acid shampoo) (1:10).

[0167] (3) Testing indicators: Measure skin moisture content (using a skin moisture meter) and TEWL (transepidermal water loss).

[0168] (4) Experimental results: The water content in the test group increased by 36% (P<0.01), the existing product control group increased by 20%, and the control group had no significant change; TEWL decreased by 25% in the test group, and decreased by 10% in the existing product control group. The above results show that the shampoo of the present invention has a good application effect and is better than the existing products.

[0169] 2. Clinical effect evaluation

[0170] (1) Experimental design: 40 volunteers (people with mild to moderate hair loss and dry hair) were recruited and randomly divided into two groups, one group used the shampoo of the present invention, and the other group used a commercial shampoo (Amino Mason amino acid shampoo). The trial period was 8 weeks. The shampoo was used 3 times a week, with a dosage of 10 mL each time.

[0171] (2) Experimental results (such as Figure 1 (shown)

[0172] 1) Anti-hair loss effect

[0173] The number of hair lost per week: the experimental group decreased by 46%, and the commercial shampoo group decreased by 22% (P<0.01).

[0174] Hair follicle density: The experimental group increased by 22%, and the commercial shampoo group increased by 10% (P<0.05).

[0175] 2) Moisturizing and repairing effect

[0176] Hair moisture content: The experimental group increased by 29%, and the commercial shampoo group increased by 18% (P<0.01).

[0177] Hair elasticity recovery rate: The experimental group increased by 28%, and the commercial shampoo group increased by 15% (P<0.05).

[0178] The above results show that the shampoo of the present invention has a good application effect, is better than the existing commercial shampoo, and has a good application prospect.

[0179] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. An amino acid shampoo, characterized in that: According to 1000g, the shampoo comprises the following components: (1) an amino acid surfactant, wherein the amino acid surfactant is an AAS1 complex: 8%-12%; (2) Polypeptide A: 0.1%-0.3%; (3) Complex plant extracts: 2%-4%; (4) Fermentation extract: 1%-2%; (5) Hyaluronic acid modified molecules: 0.3%-0.5%; (6) a thickener, wherein the thickener is xanthan gum: 0.5%-1.5%; (7) pH adjuster and deionized water: The pH adjuster is lactic acid with a pH value of 5.0-5.5, and the balance is made up of deionized water to 100%.

2. An amino acid shampoo, characterized in that: According to 1000g, the shampoo comprises the following components: (1) amino acid surfactant, wherein the amino acid surfactant is AAS1 complex: 100 g; (2) Polypeptide A: 2 g; (3) Complex plant extract: 30 g; (4) Fermentation extract: 15 g; (5) Hyaluronic acid modified molecules: 4 g; (6) thickener, wherein the thickener is xanthan gum: 10 g; (7) pH adjuster and deionized water: Use appropriate amount of lactic acid to adjust the pH to 5.0-5.5, and make up to 1000 g with deionized water.

3. The shampoo according to any one of claims 1 or 2, characterized in that: The AAS1 complex comprises the following components: (1) Sodium cocoyl glutamate: 500g; (2) Potassium cocoyl glycinate: 300 g; (3) Sodium cocoyl alanine: 100g; (4) Sodium cocoyl isethionate: 50 g; (5) Glycerol: 30 g; (6) Citric acid: Use appropriate amount of citric acid to adjust pH to 5.0-6.0; (7) Deionized water: Make up to 1000 g with deionized water.

4. The shampoo according to any one of claims 1 or 2, characterized in that: The amino acid sequence of the polypeptide A is KGPVQCGFSK, and its molecular weight is about 1175.4 Da.

5. The shampoo according to any one of claims 1 or 2, characterized in that: The composite plant extract comprises effective ingredients obtained from Polygonum multiflorum and Camellia sinensis.

6. The shampoo according to any one of claims 1 or 2, characterized in that: The total sugar content of the fermentation extract is 25.6±0.8 mg / mL; 90% of the fermentation extract is oligosaccharides, wherein the molecular weight of the oligosaccharides is <1 kDa, and the purity of the oligosaccharides is 98.3±0.4%; wherein the main components of the oligosaccharides are L-fucose, D-galactose, and mannose, and the molar ratio of L-fucose, D-galactose, and mannose is 3:2:

1.

7. The shampoo according to any one of claims 1 or 2, characterized in that: The average molecular weight of the hyaluronic acid modified molecules is ≤0.5 kDa, the distribution range is 0.3-0.5 kDa, and the proportion is ≥90%.

8. A method for preparing the shampoo according to any one of claims 1 or 2, characterized in that: The method comprises the following steps: (1) Dissolving the basic cleaning system: dissolving the AAS1 complex in 50°C deionized water and stirring for 30 minutes to form a basic cleaning solution A; (2) Active substance integration: Dissolve polypeptide A, compound plant extract and fermentation extract separately and add them to solution A one by one while stirring at a low speed; (3) Addition to the moisturizing and repairing system: Dissolve the hyaluronic acid modified molecules in a small amount of water and slowly add them to the system, keeping the temperature below 35°C; (4) Thickening and adjustment: add xanthan gum and adjust the pH to 5.0-5.5 with lactic acid, then use deionized water to make up the volume, stir evenly, filter and fill; (5) Finished product packaging: The final solution is filtered to ensure there are no bubbles, and then filled into a sterile container and sealed for storage.

9. Use of the AAS1 complex, polypeptide A, composite plant extract, fermentation extract and hyaluronic acid modified molecule as claimed in any one of claims 1 or 2 in the preparation of amino acid shampoo.

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