Anti-hair loss shampoo containing active peptide

Abstract

According to the hair loss prevention shampoo containing the active peptide, the active peptide is derived from natural plants and can promote skin cell proliferation, regulate immune factor secretion, improve the physiological environment of skin, prevent hair loss and promote hair growth, and a research basis is provided for development of related skin care products.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology research and development, and specifically provides an anti-hair loss shampoo containing active peptides. Background Art

[0002] Hair loss is a complex problem that not only affects appearance but can also cause great emotional distress. In recent years, the incidence of hair loss has been increasing year by year, seriously affecting people's physical and mental health. Compared with taking medication, people are more inclined to improve the condition of the scalp in daily hair care to inhibit and prevent hair loss. Therefore, researchers have developed a variety of shampoos with anti-hair loss functions. Shampoo is a product mainly used to clean the scalp and is usually composed of 10 to 30 ingredients, including detergents, conditioners, pH adjusters, fragrances and other additives (see Gavazzoni Dias MFR. Pro and contra of cleaning conditioners. Skin Appendage Disord. 2019; 5(03): 131–134).

[0003] Researchers have developed a variety of toiletries that can regulate the physiological functions of the scalp and inhibit hair loss. For example, Saxena et al. found that hair loss may be related to scalp bacterial and fungal disorders, and developed medicated shampoos containing ketoconazole, salicylic acid, climbazole or tar, which may help maintain a healthy scalp microbiome and treat seborrheic dermatitis (see Saxena R, Mittal P, Clavaud C. Comparison of healthy and dandruff scalp microbiome reveals the role of commensals in scalp health. Front Cell Infect Microbiol. 2018; 8: 346); hydrolyzed amino acids are protein-derived substances that can temporarily increase the strength and hydrophobicity of hair. Barba et al. used hydrolyzed amino acids from animal and plant sources as effective hair repair agents and natural moisturizers to neutralize the negatively charged network throughout the fiber and promote surface coating. They also found that the combination of hydrolyzed amino acids is more effective under heating, resulting in more stable and complete deposition on the stratum corneum cell membrane complex (see Barba C, Méndez S, Roddick-Lanzilotta A, et al. Cosmetic effectiveness of topically applied hydrolysedkeratin peptides and lipids derived from wool. Skin Res Technol. 2008; 14(02): 243–248); Plant oils have long been used to improve the lubricity and manageability of hair. Habe et al investigated the effects of mineral oil, sunflower oil, and coconut oil on preventing hair damage and concluded that coconut oil is the only oil that can reduce protein loss in damaged and undamaged hair when used as a pre- and post-shampoo agent. Coconut oil contains lauric acid, which has a high affinity for hair proteins and a low molecular weight, allowing the oil to penetrate the cortex.(See Habe T, Tanji N, Inoue S. ToF-SIMS characterization of the lipid layer on the hair surface. I: the damage caused by chemical treatments and UV radiation Surf Interface Anal. 2011; 43 (1-2): 410–412); Another common oil in hair care formulas is castor oil extracted from castor plant seeds, which is rich in castor oil, a monounsaturated fatty acid that can act as a moisturizer (see Rele AS, Mohile R B. Effect of mineral oil, sunflower oil, and coconut oil on prevention of hair damage. J Cosmet Sci. 2003; 54 (02): 175–192).

[0004] Peptides or proteins have multiple physiological activities, high bioavailability, and low side effects. Currently, many studies have used them to treat and prevent hair loss. Lee et al. found that CXXC-type zinc finger protein 5 (CXXC5) is a negative regulator of the Wnt / β-catenin pathway, and participates in hair regeneration and wound-induced hair follicle neogenesis through interaction with Disheld. CXXC5 inhibits alkaline phosphatase activity and cell proliferation, destroys the disordered interaction between CXXC5 and competitor peptides, activates the Wnt / β-catenin pathway, and accelerates hair regeneration and wound-induced hair follicle neogenesis (see Lee ST, Seo SH, Lee DH, et al. Targeting of CXXC5 by a Competing Peptide Stimulates Hair Regrowth and Wound-Induced Hair Neogenesis, J Invest Dermatol, 2017; 137(11): 2260-2269). Tsuruki et al. found that Gly-Leu-Phe (GLF) is an immunostimulating peptide derived from alpha-lactalbumin. In a neonatal rat model, intraperitoneal injection of 100 mg / kg for 4 days or oral administration of 300 mg / kg for 6 days can prevent hair loss, epidermal thickening and thinning of the fat cell layer induced by the anticancer agent etoposide (see Tsuruki T, Yoshikawa M. Anti-alopecia effect of Gly-Leu-Phe, an immunostimulating peptide derived from alpha-lactalbumin, Biosci Biotechnol Biochem. 2005; 69(8): 1633-1635). Chinese patent CN119033615A discloses an adjustable high transdermal penetration herbal composite polypeptide and its application in preventing hair loss and promoting hair growth. The liposomes encapsulating the blue copper peptide and / or the arborvitae extract are prepared by combining a core material including blue copper peptide and / or arborvitae extract and an encapsulating material including polyglycerol fatty acid ester. The liposomes have the effects of improving the expression of hair growth promoting factors, achieving good cell proliferation effect, good testosterone reduction efficiency, good oil control effect, good skin permeability, thickening hair diameter, increasing the number of hair in the growth phase, reducing the number of hair in the resting phase, and preventing hair loss.Chinese patent CN118787584A discloses a hair conditioner containing marine biological protein peptide and its preparation process, wherein the weight proportions of the components are: 5-15 parts of marine biological protein peptide, 2-6 parts of sweet almond oil, 0.1-0.3 parts of vitamin E, 0.1-0.2 parts of disodium EDTA, 0.5-1.5 parts of violet flower extract, 0.1-0.5 parts of phenoxyethanol, 0.1-0.3 parts of benzoic acid, 0.1-0.3 parts of dehydroacetic acid, 5-10 parts of cationic conditioner, 1-3 parts of emulsifier, EDTA 0.1-0.3 parts, citric acid 0.1-0.3 parts, sodium chloride 0.5-2 parts, deionized water 40-80 parts, through the marine biological hair care products, pure natural marine biological protein peptides and pod flower extract are selected as the main raw materials, and the advantages of high permeability, high diffusion, high emulsification, high surface tension, high solubility, weak alkalinity and other advantages of deionized water are utilized. While ensuring the safety of the product, these active ingredients are effectively absorbed by the scalp and hair, and effectively play an excellent hair care and anti-hair loss effect.

[0005] Although a variety of hair care products that can prevent hair loss have been developed, their effects are still unstable, and damage to hair quality and repeated symptoms often occur. For this reason, the present invention contains an anti-hair loss shampoo containing active peptides. The new active peptides are used to nourish the scalp, promote hair growth, regulate the secretion of immune factors, and effectively prevent hair loss. Summary of the invention

[0006] The first aspect of the present invention provides an active peptide, the amino acid sequence of which is shown in SEQ ID NO: 1. The active peptide is derived from natural plant extracts, has strong biological activity, can promote skin cell proliferation and hair growth, and regulate the secretion of immune factors.

[0007] The second aspect of the present invention provides a shampoo comprising the active peptide.

[0008] Furthermore, the shampoo comprises, by mass percentage, 1%-3% of the active peptide, 10%-20% of a surfactant, 0.5%-5% of a thickener, 0.5-1% of a preservative, 2%-10% of sodium hyaluronate, 1%-5% of a polyquaternium salt, and the remainder is water.

[0009] Further, it comprises 2% of the active peptide.

[0010] Furthermore, the surfactant is one or more of sodium lauryl sulfate, sodium dodecylbenzene sulfate and cocoyl glyceryl betaine.

[0011] Furthermore, the surfactant is sodium lauryl sulfate.

[0012] Furthermore, the thickener is one or more of glycerin, propylene glycol, xanthan gum, guar gum and ceteareth-60 myristyl glycol.

[0013] Furthermore, the thickener is glycerol.

[0014] Furthermore, the preservative is one or more of phenoxyethanol, ethylhexylglycerin, 1,2-hexanediol, and 1,2-pentanediol.

[0015] Furthermore, the preservative is 1,2-hexanediol.

[0016] Beneficial Effects

[0017] The present invention provides an anti-hair loss shampoo containing active peptides, which has the following beneficial effects:

[0018] 1. New active peptides were screened and obtained from natural plant extracts, which can promote skin cell proliferation;

[0019] 2. The active peptide can effectively promote hair growth;

[0020] 3. The active peptide can regulate the secretion of immune factors, improve the physiological environment of the skin, and prevent hair loss;

[0021] 4. The shampoo containing the active peptide can effectively maintain the function of the active ingredients and can be used for skin care. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 : Active peptides promote the proliferation of skin fibroblasts;

[0023] Figure 2 :HE staining of skin tissue sections;

[0024] Figure 3 : VEGF expression level in skin tissue;

[0025] Figure 4 : Expression level of TGF-β in skin tissue. DETAILED DESCRIPTION

[0026] The experimental methods described in the following examples are conventional methods unless otherwise specified; the reagents, biological materials, and detection kits are all commercially available unless otherwise specified.

[0027] Example 1 Screening and preparation of ginger active peptides

[0028] Ginger is a common medicinal plant, also known as ginger root, Bai La Yun, Yin Di Xin, spicy taste, slightly warm in nature, with the effects of dispelling cold, warming the middle and stopping vomiting, warming the lungs and stopping cough, and detoxification. Chinese researchers have found that ginger has the function of promoting hair follicle growth and preventing hair loss (see He Jingyu, Chen Qingsheng, Gong Shengzhao, etc., Extraction and separation of active substances from ginger plants with anti-hair loss effect and structure-activity relationship research and industrialization. Science and Technology Achievements, 2021; Liu Qing, Huang Fang, Anti-hair loss effect of composite extracts of Platycladus orientalis, ginger, red clover, and mugwort. Guangdong Chemical Industry. 2022; 49(22): 76-78). In the present invention, ginger extract is used as a raw material to obtain ginger protein therein, and then alkaline protease is used for hydrolysis to separate and obtain short peptides with biological activity, including 6 short peptides with high activity, which are named GG01, GG02, GG03, GG04, GG05 and GG06 respectively.

[0029] In order to further study the promoting effect of the active peptide on skin and hair growth, this section uses human skin fibroblasts BJ-5ta as the object to examine the ability of the above-mentioned activity to promote cell proliferation. BJ-5ta cells were cultured at 37°C and 5% CO2. When they grew to a density of 80%-90%, the cells were digested and collected by centrifugation. 4 / The cell density was added to the 96-well plate, 100 μL per well. After overnight culture, the cells were attached to the wall, and 50 ng / mL of GG01-GG06 active peptides were added respectively. The experimental wells with equal amount of culture medium were used as negative controls, and the experimental wells without cells were used as blank controls. After 24 hours of co-culture, 10 μL CCK-8 reagent was added to each well, and the cells were incubated at 37 ° C for 4 hours. After the color turned yellow, the OD value of each group was measured by the microplate reader. The OD values ​​of the experimental group were compared with those of the control group, and the cell proliferation rate of each group was calculated.

[0030] Cell proliferation rate = (average OD value of experimental group - average OD value of blank control group) / (average OD value of negative group - average OD value of blank control group) × 100%

[0031] The results are as follows Figure 1 As shown, the above active peptides have different degrees of promoting proliferation of fibroblasts, which is beneficial to improving the physiological environment of the skin. Among them, GG05 has the strongest promoting effect and is the candidate active substance for subsequent experiments. After identification, the amino acid sequence of GG05 is shown in SEQ ID NO:1.

[0032] According to the amino acid sequence structure of GG05, it was prepared by solid phase synthesis and then purified and freeze-dried for subsequent experiments.

[0033] Example 2 Preparation and administration of mouse hair loss model

[0034] Androgenic alopecia is the most common type of hair loss. According to incomplete statistics, about 90% of hair loss belongs to this type. Therefore, the present invention is based on the androgenic alopecia animal model to study the preventive and therapeutic effects of ginger active peptide on animals.

[0035] C57BL / 6 mice were adaptively fed for 7 days and randomly divided into 3 groups, 10 mice in each group, namely normal group, active peptide group and control group. In the normal group, only the back hair was removed without modeling; in the other two groups, after the back hair was removed, 0.1 mg of testosterone propionate was injected into the back at multiple points every day for 20 days for modeling. In addition, the active peptide group was injected with 50 mg / kg GG05 active peptide at multiple points on the back every day for 30 days; the control group was injected with an equal amount of saline at multiple points on the back every day for 30 days.

[0036] Example 3 Skin Histological Observation

[0037] On the second day after stopping the drug administration, mice in each group were killed by cervical dislocation, and the back skin was taken and fixed with 4% paraformaldehyde. After paraffin embedding, skin sections were prepared, fixed and sealed after HE staining, and observed under a microscope. Figure 2 As shown, in the normal group, the number of hair follicles was large, the distribution was uniform, the hair was thick and in good growth condition; in the control group, after modeling treatment, the number of hair follicles was greatly reduced, the distribution was disordered, and normal hair growth was not observed; in the active peptide group, after treatment, the number of hair follicles increased, the arrangement was dense, and the hair diameter became thicker, indicating that it has a good promoting effect on hair growth.

[0038] Example 4 Immune Factor Secretion

[0039] The skin of the experimental area of ​​mice was taken, the skin tissue was cut into small pieces, and the tissue lysis buffer was added at a mass volume ratio of 1:9, and the tissue was fully ground into a homogenate at 4°C, centrifuged at 3000r / min for 10min, and the supernatant was collected. The ELISA kit (purchased from Shanghai Xitang Biotechnology Co., Ltd.) was used to detect the VEGF and TGF-β content in the supernatant. The specific steps were carried out according to the instructions of the kit.

[0040] The results are as follows Figure 3 , Figure 4 As shown in the figure, compared with the normal group, the VEGF expression level was significantly reduced after modeling, while the TGF-β expression level was significantly increased. After active peptide treatment, the VEGF expression level increased and was close to the normal level; while the expression of TGF-β was inhibited, and although it was still higher than the normal level, it was significantly lower than the control group.

[0041] Example 5 Preparation of Shampoo Containing Active Peptides

[0042] The shampoo provided in the present invention comprises, by mass percentage, 1%-3% of GG05 active peptide, 10%-20% of surfactant, 0.5%-5% of thickener, 0.5-1% of preservative, 2%-10% of sodium hyaluronate, 1%-5% of polyquaternium salt, and the rest is water. Among them, the surfactant is one or more of sodium lauryl sulfate, sodium dodecylbenzene sulfate and cocoyl gum propyl betaine, the thickener is one or more of glycerin, propylene glycol, xanthan gum, guar gum and ceteareth-60 myristyl glycol, and the preservative is one or more of phenoxyethanol, ethylhexylglycerin, 1,2-hexanediol and 1,2-pentanediol. As a preferred technical solution, the shampoo includes 2% GG05 active peptide, 10% surfactant, 3% thickener, 0.5% preservative, 5% sodium hyaluronate, 1% polyquaternium salt, and the rest is water; the surfactant is preferably sodium lauryl sulfate, the thickener is preferably glycerol, and the preservative is preferably 1,2-hexanediol.

[0043] The preparation method of the shampoo is as follows: weighing various raw materials according to mass percentage; heating water to about 50° C., adding a surfactant and mixing thoroughly; then adding GG05 active peptide, a thickener, a preservative, sodium hyaluronate and a polyquaternium salt, stirring thoroughly for 20-30 minutes, and cooling to obtain the shampoo.

[0044] After testing, the shampoo can remain stable at room temperature and can be stored for more than 6 months and still has anti-hair loss activity.

[0045] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that the technical solutions described in the above embodiments may still be modified, or some or all of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. An active peptide, characterized in that The amino acid sequence of the active peptide is shown in SEQ ID NO:

1.

2. A shampoo, characterized in that: The invention comprises the active peptide described in claim 1.

3. The shampoo according to claim 2, characterized in that The shampoo comprises, by mass percentage, 1%-3% of the active peptide, 10%-20% of a surfactant, 0.5%-5% of a thickener, 0.5-1% of a preservative, 2%-10% of sodium hyaluronate, 1%-5% of a polyquaternium salt, and the remainder is water.

4. The shampoo according to claim 3, characterized in that 2% of the active peptide was included.

5. The shampoo according to claim 3, characterized in that The surfactant is one or more of sodium lauryl sulfate, sodium dodecylbenzene sulfate and cocoyl gum propyl betaine.

6. The shampoo according to claim 5, characterized in that The surfactant is sodium lauryl sulfate.

7. The shampoo according to claim 3, characterized in that The thickener is one or more of glycerin, propylene glycol, xanthan gum, guar gum and ceteareth-60 myristyl glycol.

8. The shampoo according to claim 7, characterized in that The thickener is glycerin.

9. The shampoo according to claim 3, characterized in that Preservatives are phenoxyethanol, ethylhexylglycerin, One or more of 1,2-hexanediol and 1,2-pentanediol.

10. The shampoo according to claim 7, characterized in that The preservative is 1,2-hexanediol.

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