Application of bioactive peptide extracted from human hair in preparation of anti-alopecia and hair-growing medicine

Abstract

The invention discloses an application of a human hair extracted bioactive peptide in preparation of an anti-alopecia and hair growth medicament, and the anti-alopecia and hair growth medicament. The bioactive peptide extracted from human hair has very high biological activity, induces and promotes proliferation and differentiation of hair follicle cells, and promotes hair growth; the shampoo has the effects of expanding scalp capillary vessels, improving scalp blood microcirculation, activating hair growth hair cells, shortening the abnormal resting period of hair, prolonging the growth period of hair, effectively preventing alopecia and promoting hair growth. Through standard test and statistical analysis, the anti-hair loss counting result is as follows: after 12 weeks of use, the anti-hair loss counting result is compared with the basic value before use, and p is less than 0.05. According to the hair growth counting result, after 18 weeks of use, the basic value is compared with the basic value before use, and p is smaller than 0.01. The bioactive peptide extracted from human hair is a natural raw material, is good in safety, strong in controllability and stable in performance, has multiple benefits and activity diversity, and meets the requirements of different products.

Description

Technical Field

[0001] The invention specifically relates to application of a bioactive peptide extracted from human hair in preparing a hair loss prevention and hair growth medicine. Background Art

[0002] With the increase of life pressure and the influence of environmental factors, the problem of hair loss is becoming more and more serious; especially among young people, the phenomenon of hair loss is becoming more and more common. Whether it is men or women, hair loss not only affects appearance, but may also have a negative impact on mental health. According to statistics, about one-third of adults will experience different degrees of hair loss in their lifetime. Consumers' demand for anti-hair loss and hair growth products is growing. This trend has promoted the rapid development of the anti-hair loss and hair growth product market, and many anti-hair loss and hair growth products have emerged in the market. The popular anti-hair loss and hair growth products on the market currently include minoxidil, finasteride, dutasteride, bimatoprost, KOSE hair care products, spironolactone, lipid-lowering hair growth tablets, scalp care products (such as shampoo, essence), etc.

[0003] The main ingredients of the first four products mentioned above are all made through chemical synthesis; their advantages are: ① strong controllability, the molecular structure of chemically synthesized ingredients can be precisely controlled to ensure their efficacy and stability; ② good cost-effectiveness, in large-scale production, chemically synthesized raw materials are usually cheaper than natural extracts; ③ strong versatility, a variety of different active ingredients can be synthesized to meet the needs of different products; their disadvantages are: ① safety issues, some synthetic ingredients may cause skin irritation or allergic reactions; ② environmental impact, the chemical synthesis process may cause pollution to the environment.

[0004] The last four products (series) mentioned above are all products that combine traditional herbal essences, specifically by extracting bioactive ingredients and nutrients from a variety of Chinese herbal medicines and natural plants; their advantages are: ① naturalness, generally considered safer, with fewer allergic reactions, and suitable for long-term use; ② multiple benefits, many plant extracts have multiple biological activities, can improve the health of hair and scalp at the same time, suitable for a variety of hair loss types and various hair qualities, and feel good to use; ③ easy to use, suitable for daily care; ④ cultural identity, in some markets, consumers prefer to use traditional Chinese herbal products; their disadvantages are: ① unstable ingredients, the active ingredients of natural extracts contain ① The amount may vary due to factors such as season and origin; ② The effect is slow and may not be as obvious as that of drugs. Some users may need to use it continuously to see the effect; ③ There are obvious individual differences, and some users may be sensitive to certain ingredients; ④ The extraction of functional ingredients for anti-hair loss and hair growth from traditional Chinese herbal medicine and natural plants requires the use of organic solvents, mostly ethanol; and long-term use of products containing ethanol will have adverse effects on scalp health, among which dryness, irritation, inflammation and hair health problems are all potential risks, and may also lead to an imbalance in the oil secretion of the scalp, thereby inducing seborrheic dermatitis; ⑤ The extraction process of some plants is complicated and costly. Summary of the invention

[0005] The purpose of the present invention is to provide an application of a bioactive peptide extracted from human hair in the preparation of anti-hair loss and hair growth medicines.

[0006] Another object of the present invention is to provide a hair loss prevention and hair growth medicine with good efficacy and high safety.

[0007] The purpose of the present invention is achieved through the following measures: application of bioactive peptides extracted from human hair in the preparation of anti-hair loss and hair growth drugs.

[0008] Another object of the present invention is achieved by the following measures: an anti-hair loss and hair growth medicine comprising bioactive peptides extracted from human hair.

[0009] The human hair extracted bioactive peptides of the present invention are products obtained by precisely controlling the hydrolysis conditions to achieve the goal of limited hydrolysis (incomplete hydrolysis) during the hydrolysis of human hair, that is, the peptide segments that do not express biological activity in human hair proteins can be released to the maximum extent to express biological activity, while the amino acid sequences of the biologically active peptide segments that can already be expressed are not excessively hydrolyzed and destroyed; these fixed amino acid sequence peptide segments with biological activity are called bioactive peptides (BAPs), which mainly contain a mixture of multiple biologically active peptides with a molecular weight of 300 to 7000D.

[0010] The preparation method of the bioactive peptide extracted from human hair of the present invention is disclosed in the invention patent CN103265615A, and the specific preparation method comprises the following steps:

[0011] (1) Weigh human hair that has been washed, defatted and dried, add 7 to 10 times 0.4 to 2.0 mol / L disulfide bond reducing agent, and treat at 70 to 100° C. for 0.3 to 1 hour; remove the remaining disulfide bond reducing agent in the reaction product by dialysis, and dry at 80±10° C.;

[0012] (2) adding 7 to 10 times 5 to 10 mol / L NH2CONH2 to the product obtained in step (1), treating at 30 to 80°C for 12 to 24 hours; removing the remaining NH2CONH2 in the product after the reaction by dialysis, and drying at 80±10°C;

[0013] (3) adding 7 to 10 times 0.2 to 1 mol / L CH5N3·HCl to the product obtained in step (2), and treating at 30 to 80°C for 0.5 to 2 hours; removing the remaining CH5N3·HCl in the product after the reaction by dialysis, and drying at 80±10°C;

[0014] (4) adding 7 to 10 times of 0.5 to 3.0 mol / L NaOH to the product obtained in step (3), treating at 30 to 100° C. for 0.5 to 4 hours, and adjusting the pH to 7.3 to 7.5 with HCl;

[0015] (5) diluting or concentrating the product obtained in step (4), adjusting the BAPs content to 40-50 mg / mL by spectrophotometry, adjusting the osmotic pressure to 300±20 mOsm / L by dialysis or adding NaCl using an osmometer, and packaging and sterilizing to obtain a liquid formulation of BAPs;

[0016] Alternatively, the product obtained in step (4) is diluted or concentrated, the BAPs content is adjusted to 90-100 mg / mL by spectrophotometry, the osmotic pressure is adjusted to 60±10 mOsm / L by dialysis or addition of NaCl using an osmometer, freeze-dried, packaged and sterilized to obtain a solid-phase dosage form of BAPs.

[0017] The human hair extracted bioactive peptides of the present invention have high biological activity, induce and promote the proliferation and differentiation of hair follicle cells, promote hair growth; dilate scalp capillaries, improve scalp blood microcirculation, activate hair matrix cells for hair growth, shorten the abnormal resting period of hair, prolong the growth period of hair, effectively prevent hair loss and promote hair growth. After standardized testing and statistical analysis, the anti-hair loss count results: compared with the basic value before use after 12 weeks of use, p < 0.05; the hair growth count results: compared with the basic value before use after 18 weeks of use, p < 0.01.

[0018] In the prior art, the sources of raw materials for anti-hair loss and hair growth products mainly include three aspects: 1. chemical synthesis, 2. traditional Chinese herbal medicine and natural plant extraction, and 3. biotechnology extraction. The human hair extracted bioactive peptides described in the present invention, as raw materials for anti-hair loss and hair growth drugs, do not come from these three aspects, but are obtained by hydrolyzing human hair in limited amounts using a hydrolysis method.

[0019] The human hair extracted bioactive peptides have the advantages of the raw material products from the three sources mentioned above, while avoiding their disadvantages; and also have the following advantages: ① natural raw materials, good safety, and very few allergic reactions; ② the product is highly controllable, the hydrolysis conditions are controllable, and there is no instability in the active ingredient content of natural extracts that may vary due to factors such as seasons and origins; ③ it is widely applicable and can hydrolyze a variety of peptides with different biological activities to meet the needs of different products; ④ hydrolysis does not require the use of organic solvents, eliminating its adverse effects and potential risks such as scalp irritation; ⑤ the technical threshold is not high, and does not require a high level of technology and equipment investment; ⑥ it is easy to scale up production and is cost-effective; ⑦ the preparation method does not cause environmental impact. DETAILED DESCRIPTION

[0020] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. 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 creative work are within the scope of protection of the present invention.

[0021] Embodiment 1

[0022] (1) Quantitatively weigh washed, degreased and dried human hair and add 7 times the mass of 1.0 mol / L C4H 10 O2S2, treated at 90℃ for 0.7 hours; the remaining C4H 10 O2S2, dried at 80±10℃;

[0023] (2) adding 8 mol / L NH2CONH2 in an amount of 9 times the mass to the product obtained in step (1), and treating at 60°C for 18 hours; removing the remaining NH2CONH2 in the product after the reaction by dialysis, and drying at 80±10°C;

[0024] (3) adding 9 times the mass of 0.6 mol / L CH5N3·HCl to the product obtained in step (2), and treating at 50°C for 1.0 hour; removing the remaining CH5N3·HCl in the product after the reaction by dialysis, and drying at 80±10°C;

[0025] (4) Add 8 times the mass of 1.0 mol / L NaOH to the product obtained in step (3), treat at 90° C. for 1.0 hour, and adjust the pH to 7.4 with HCl; adjust the BAPs content to 50 mg / mL by dilution and spectrophotometry, adjust the osmotic pressure to 300±20 mOsm / L by adding NaCl and using an osmometer, and package and sterilize to obtain a liquid formulation of BAPs.

[0026] The prepared liquid formulation BAPs is used as an anti-hair loss and hair growth drug. The product has stable performance and can be sterilized with pressure steam to give a sterile shelf life of up to 3 years. It is also easy to store and transport. The raw material source is sufficient, and it is easy to form large-scale production at a low cost.

[0027] Embodiment 2

[0028] (1) Quantitatively weigh human hair that has been washed, defatted, and dried, add 8 times the mass of 2.0 mol / L C2H6OS, and treat at 80°C for 0.8 h; remove the remaining C2H6OS in the reaction product by dialysis, and dry at 80±10°C;

[0029] (2) adding 10 times the mass of 8 mol / L NH2CONH2 to the product obtained in step (1), and treating at 70°C for 10 hours; removing the remaining NH2CONH2 in the product after the reaction by dialysis, and drying at 80±10°C;

[0030] (3) adding 8 times the mass of 0.9 mol / L CH5N3·HCl to the product obtained in step (2), and treating at 60°C for 1.5 hours; removing the remaining CH5N3·HCl in the product after the reaction by dialysis, and drying at 80±10°C;

[0031] (4) Add 9 times the mass of 2.4 mol / L NaOH to the product obtained in step (3), treat at 50°C for 3 hours, and adjust the pH to 7.4 with HCl; after concentration, adjust the BAPs content to 100 mg / mL by spectrophotometry. After dialysis, adjust the osmotic pressure to 60±10 mOsm / L with an osmometer.

[0032] (5) The product obtained in step (4) is freeze-dried, packaged and sterilized to prepare a solid-phase dosage form BAPs.

[0033] The prepared solid-phase dosage form BAPs is used as an anti-hair loss and hair growth drug. The product has stable dosage performance and can be sterilized with pressure steam to have a sterile shelf life of up to 2 years. It is easy to store and transport. The raw material source is sufficient, and it is easy to form large-scale production with low cost.

[0034] Embodiment 3

[0035] Experimental evaluation of the anti-hair loss efficacy of the liquid BAPs prepared in Example 1 on human body.

[0036] The test product is the liquid BAPs and matrix formula product prepared in Example 1, and the control product is the matrix formula product without the liquid BAPs prepared in Example 1. The product in the washout period is the same as the control product, and both the test group and the control group use the control product during the washout period and the test period.

[0037] A total of 30 subjects completed the test, aged 21 to 53 years old. Among them, 20 subjects were in the experimental group, all of whom were female, with an average age of 35±8.09 ​​years old; 10 subjects were in the control group, including 1 male and 9 females, with an average age of 36±5.66 years old. There was no statistical difference in gender and age between the two groups (P<0.05), and the subjects met the voluntary inclusion and exclusion criteria.

[0038] 1. Test methods:

[0039] The test was carried out in accordance with the specific requirements of the technical specification method "Test Method for Anti-Hair Loss Efficacy of Cosmetics" released in February 2021. The qualified subjects enrolled in the group first used the wash-out period products for two weeks. After the wash-out period, the subjects who met the requirements were divided into the test group and the control group according to the random table. The test products and control products were distributed respectively, and the products were used correctly for 12 weeks according to the product instructions. During the trial, the subjects were required to keep a usage diary and strictly abide by the restrictions. The visits were conducted before and 4 weeks, 8 weeks and 12 weeks after the use of the product, and hair loss counts, hair density assessment tests and image photography were performed at each visit.

[0040] 2. Test results:

[0041] Table 1 Hair loss counting results before and after product use

[0042]

[0043] * Statistical analysis was performed using the rank sum test, compared with the baseline value before use, p < 0.05.

[0044] Table 2 Difference in hair loss counts before and after product use

[0045] Test object 4 weeks 8 weeks 12 weeks Test products -4.10±9.80 -5.15±11.47 -6.55±10.65 Comparison products -6.80±9.07 -4.40±7.47 -4.70±11.03

[0046] Table 3 Overall hair density evaluation results before and after product use [median (minimum, maximum)]

[0047]

[0048] * Statistical analysis was performed using the rank sum test, compared with the baseline value before use, p < 0.05.

[0049] Table 4 Difference in overall hair density before and after product use [median (minimum, maximum)]

[0050]

[0051] * Statistical analysis was performed using the rank sum test, compared with the baseline value before use, p < 0.05.

[0052] Table 5 Results of local hair quantity before and after product use

[0053]

[0054] Table 6 Difference in the number of local hairs before and after using the product

[0055] Test object 4 weeks 8 weeks 12 weeks Test products -0.74±13.53 3.80±10.39 -0.67±14.41 Comparison products 3.20±13.96 2.10±13.66 2.00±12.57

[0056] Table 7 Results of local hair density before and after product use

[0057]

[0058] Table 8 Difference in local hair density before and after product use

[0059] Test object 4 weeks 8 weeks 12 weeks Test products -0.82±14.97 4.19±11.50 -0.74±15.96 Comparison products 3.54±15.45 2.33±15.11 2.15±13.91

[0060] 3. Test conclusion:

[0061] The results of the anti-hair loss efficacy test of the liquid BAPs prepared in Example 1 showed that the hair loss count results after 12 weeks of use were compared with the baseline value before use, p < 0.05. The visual score and difference of the overall hair density, image score, and the mean and difference of the overall hair density score of the test product after 12 weeks of use were compared with the baseline value before use, p < 0.05. In summary, the product has an anti-hair loss efficacy.

[0062] Embodiment 4

[0063] The solid phase BAPs prepared in Example 2 were tested and observed to evaluate the hair growth effect on human body.

[0064] Qualified healthy volunteers were selected according to the inclusion and exclusion criteria, and the formula matrix product was used according to the instructions for use for 2 weeks. A total of 30 effective subjects completed the test, aged 24 to 56 years old, and were randomly divided into an experimental group and a control group. Among them, 20 subjects were in the experimental group, all male, with an average age of 37±6.9 years old; 10 subjects were in the control group, 9 males and 1 female, with an average age of 36±5.1 years old. There was no statistical difference in gender and age between the two groups (P<0.05), which met the voluntary inclusion and exclusion criteria of the subjects.

[0065] The test product was a formula matrix containing 5% solid phase BAPs, and the control product was a formula matrix without solid phase BAPs. The test product and control product were distributed to the test group and the control group, respectively, and all participants knew that they were using the test product. The product was used correctly according to the product instructions for 18 weeks. During the trial, the subjects were required to keep a usage diary and strictly abide by the restrictions.

[0066] The patients were followed up before and 6, 12, and 18 weeks after product use. A hair densitometer was used to count the number of hairs per square centimeter in a specific area, and images were taken.

[0067] Table 10 Hair count results before and after using solid phase BAPs products

[0068]

[0069]

[0070] * Statistical analysis was performed using the rank sum test, compared with the baseline value before use, p < 0.05.

[0071] ** Statistical analysis was performed using the rank sum test, compared with the baseline value before use, p < 0.01.

[0072] The results of the hair growth efficacy test of the solid phase BAPs prepared in Example 2 showed that the hair growth count results after 12 weeks of use were compared with the baseline value before use, p < 0.05. After 18 weeks of use, compared with the baseline value before use, p < 0.01, indicating that the product has a very significant hair growth efficacy.

Claims

1. Application of bioactive peptides extracted from human hair in the preparation of anti-hair loss and hair growth drugs; The human hair extracted bioactive peptides are prepared by the following steps: (1) Weigh human hair that has been washed, defatted and dried, add 7 to 10 times 0.4 to 2.0 mol / L disulfide bond reducing agent, and treat at 70 to 100° C. for 0.3 to 1 hour; remove the remaining disulfide bond reducing agent in the reaction product by dialysis, and dry at 80±10° C.; (2) adding 7 to 10 times 5 to 10 mol / L NH2CONH2 to the product obtained in step (1), treating at 30 to 80°C for 12 to 24 hours; removing the remaining NH2CONH2 in the product after the reaction by dialysis, and drying at 80±10°C; (3) adding 7 to 10 times 0.2 to 1 mol / L CH5N3·HCl to the product obtained in step (2), and treating at 30 to 80°C for 0.5 to 2 hours; removing the remaining CH5N3·HCl in the product after the reaction by dialysis, and drying at 80±10°C; (4) adding 7 to 10 times of 0.5 to 3.0 mol / L NaOH to the product obtained in step (3), treating at 30 to 100° C. for 0.5 to 4 hours, and adjusting the pH to 7.3 to 7.5 with HCl; (5) diluting or concentrating the product obtained in step (4), adjusting the BAPs content to 40-50 mg / mL by spectrophotometry, adjusting the osmotic pressure to 300±20 mOsm / L by dialysis or adding NaCl using an osmometer, and packaging and sterilizing to obtain a liquid formulation of BAPs; Alternatively, the human hair extracted bioactive peptides are prepared by the following steps: (1) Weigh human hair that has been washed, defatted and dried, add 7 to 10 times 0.4 to 2.0 mol / L disulfide bond reducing agent, and treat at 70 to 100° C. for 0.3 to 1 hour; remove the remaining disulfide bond reducing agent in the reaction product by dialysis, and dry at 80±10° C.; (2) adding 7 to 10 times 5 to 10 mol / L NH2CONH2 to the product obtained in step (1), treating at 30 to 80°C for 12 to 24 hours; removing the remaining NH2CONH2 in the product after the reaction by dialysis, and drying at 80±10°C; (3) adding 7 to 10 times 0.2 to 1 mol / L CH5N3·HCl to the product obtained in step (2), and treating at 30 to 80°C for 0.5 to 2 hours; removing the remaining CH5N3·HCl in the product after the reaction by dialysis, and drying at 80±10°C; (4) adding 7 to 10 times of 0.5 to 3.0 mol / L NaOH to the product obtained in step (3), treating at 30 to 100° C. for 0.5 to 4 hours, and adjusting the pH to 7.3 to 7.5 with HCl; (5) The product obtained in step (4) is diluted or concentrated, and the BAPs content is adjusted to 90-100 mg / mL by spectrophotometry. The osmotic pressure is adjusted to 60±10 mOsm / L by dialysis or by adding NaCl using an osmometer, and then freeze-dried, packaged and sterilized to obtain a solid-phase dosage form of BAPs.

2. A hair loss prevention and hair growth medicine comprising the bioactive peptide extracted from human hair according to claim 1.

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