Hair growth cosmetic and preparation method thereof

Abstract

The invention relates to a hair growth cosmetic and a preparation method thereof, in particular to a cyclic dipeptide compound with a hair growth promoting effect, a cosmetic containing the compound and a preparation process of the cosmetic. The ring (Trp-Arg) and the cosmetic containing the ring (Trp-Arg) are applied through skin (especially head skin) and have a good hair growth promoting effect, compared with a control group, the hair growth effect of the ring (Trp-Arg) and the cosmetic containing the ring (Trp-Arg) is improved along with increase of the concentration, and when the concentration reaches 2%, the effect of promoting proliferation of human outer hair root sheath cells can reach about 111.8%. The proliferation promotion effects of the traditional Chinese medicine composition on human hair papilla cells and human epidermal keratinocytes can respectively reach 112.4% and 146.6%. The invention further prepares shampoo, hair conditioner and hair cream containing the rings (Trp-Arg), and model animal experiments further prove that the cosmetics containing the rings (Trp-Arg) can effectively promote the growth of the hair, especially the growth of the hair.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a cyclic dipeptide compound that promotes hair growth, as well as cosmetics containing the compound and their preparation process. Background Technology

[0002] A normal person's scalp has approximately 100,000 to 150,000 hair follicles, while Asians typically have around 100,000. Losing fewer than 100 hairs per day is normal, and the average person's hair grows about 1 centimeter per month.

[0003] The growth cycle of each hair follicle—how long it grows and when it falls out—is determined by genes, but is also influenced by many environmental factors. These include damage from dyeing and perming, overly stimulating shampoos, scalp microbial infections, irregular lifestyles, overwork, air pollution, hormonal imbalances, poor dietary habits, and excessively strong flavors.

[0004] Hair growth occurs in three phases during the hair follicle's growth cycle: telogen (resting phase), anagen (growth phase), and catagen (transitional phase). In each phase, the hair follicle responds differently to various stimuli, thus affecting the visible appearance of the hair. During the telogen phase, the hair follicle is almost at rest, located in the dermis. The hair bulb structure at the base of the follicle disappears, exposing the dermal papilla, where the dermal papilla cells are tightly packed and spherical. When the hair follicle receives a signal and begins to transition from the telogen phase to the anagen phase, stem cells located in the follicle bulge become activated, stimulating specific cells beneath them to divide and begin to envelop the enlarged dermal papilla, forming the hair bulb structure and growing downwards. Once the hair follicle enters the anagen (growth) phase, the dermal papillae surrounding the hair follicle bulb stimulate cell division in the surrounding cells. These cells then receive various signals and begin to differentiate into the multi-layered structure within the hair follicle, such as the outer root sheath, inner root sheath, matrix cells, and hair shaft. Melanin also gradually begins to be produced. At this time, the hair follicle bulb grows downwards from the dermis into the subcutaneous tissue, and the hair shaft also lengthens, emerging from the skin surface to form a visible hair. Towards the end of the anagen phase, the hair prepares to enter the telogen (resting) phase, ceasing cell division. After the hair bulb shrinks and enters the telogen phase, the hair will fall out when gently combed. After the telogen phase ends, new fine hairs begin to grow, entering the next anagen phase.

[0005] Losing more than 100 hairs a day may be due to some abnormal phenomenon causing increased hair loss. Known common causes are as follows:

[0006] ·menopause

[0007] Male pattern baldness

[0008] Long-term stress

[0009] Long-term sleep deprivation

[0010] Postpartum hair loss

[0011] Hair loss in women

[0012] ● Hair loss during the resting period

[0013] ● Iron deficiency anemia

[0014] ●Hypothyroidism

[0015] ●Circular bald patches

[0016] Folliculitis can be caused by prolonged and repeated exposure to irritants.

[0017] Less common causes of hair loss include pulling on the hair, using certain medications such as chemotherapy, HIV / AIDS, hypothyroidism, and malnutrition. For example, iron and protein deficiencies can cause hair to turn yellow and split; deficiencies in vegetable oils, vitamin A, protein, and iodine can cause hair to become dry, dull, and brittle; and vitamin B deficiencies can lead to seborrheic dermatitis and hair loss. Causes of hair loss that manifest as inflammation or scarring include fungal infections, lupus erythematosus, radiation therapy, and sarcoidosis.

[0018] Hair loss is a very common problem. Among people aged 50, roughly half of men and a quarter of women are affected; and about 2% will start to experience alopecia areata at some point.

[0019] Traditional hair growth products, such as those containing glyceryl pentadecanoate, adenosine, minoxidil, plant-based caffeine extract, and saw palmetto extract as their main components, primarily focus on replenishing nutrients, promoting blood circulation, and improving scalp cell activation.

[0020] Adenosine and minoxidil are known to act on dermal papilla cells, inducing the production of cell growth factors and activating hair follicle cells. However, these hair growth agents have not provided truly satisfactory results. In recent years, with technological advancements, people's expectations for effective hair growth agents have been steadily increasing.

[0021] Cyclic dipeptides, also known as 2,5-dioxopiperazines or 2,5-diketopiperazines, are formed by the cyclic linkage of two amino acids and are the smallest cyclic peptides found in nature. Naturally occurring cyclic dipeptides generally consist of a relatively stable six-membered ring formed by an α-amino acid (often an L-amino acid). Many cyclic dipeptides are endogenous in plants and animals, while others are formed through fermentation or during food processing.

[0022] Guillermo Schmeda-Hirschmann et al. reported a cyclic dipeptide from the Chilean hazelnut cotyledons (Gevuina avellana Mol, Proteaceae), Scientific Reports (2020) 10:7070. In the literature, the authors isolated a cyclic dipeptide compound from Chilean hazelnut leaves, namely cyclic tryptophan-arginine (cyclic (Trp-Arg)), CAS number: 83481-40-07, relative molecular mass: 342.4, and its chemical structure is as follows:

[0023]

[0024] The aforementioned literature also provides a method for isolating cyclic tryptophan arginine (cyclic (Trp-Arg)), but does not provide any use for this dipeptide.

[0025] A search revealed that some of the applications of the above-mentioned compounds have been reported in existing literature.

[0026] Rozenn Trepos et al., Antifouling Compounds from the Sub-Arctic Ascidian

[0027] Synoicum pulmonaria:Synoxazolidinones A and C,Pulmonarins A and B,and

[0028] Synthetic Analogues, J.Nat.Prod. 2014, 77, 2105-2113. The above-mentioned compound (compound 8) is reported to be an active inhibitor of bacterial adhesion and growth (antibacterial activity). It has a weak effect on microalgae and no effect on barnacles.

[0029] SHINOBU SAKURADANI et al., in their study "Antinociceptive Activities of Synthetic Dipeptides in Mice," indicated that cyclic tryptophan-arginine possesses analgesic activity, and that after intraventricular injection in mice, the half-maximal effective concentration (ED50) was [value missing]. 50 The average value is 41.0 nmol / animal.

[0030] Chinese patent CN106456633A indicates that cyclic tryptophan arginine (compound number 189) has GLP-1 agonist activity and can regulate human glucose metabolism.

[0031] The aforementioned dipeptides can be conveniently synthesized using conventional processes, for example, referring to Cook et al., Efficient one-step synthesis of diastereoisomeric cyclic dipeptides from amino acids: three diastereoisomers of cyclo-L-isoleucyl-L-isoleucine, J. Chem. Soc., Perkin Trans. 1, 1992, 1199-1201.

[0032] L-tryptophan and L-arginine were dissolved in anhydrous ethylene glycol, heated under reflux for 24 hours, and cooled to precipitate a solid. The solid was recrystallized in ethanol or separated by silica gel column chromatography to obtain a cyclic dipeptide. Summary of the Invention

[0033] Given the strong demand for hair care and growth products among the general public, especially men who care about their appearance, the applicant conducted in-depth research on existing peptides that may promote hair growth. Unexpectedly, they discovered that Trp-Arg has a significant effect on promoting hair growth and can be administered through the scalp.

[0034] The Trp-Arg ring is highly water-soluble, mild, non-irritating, and has good safety for topical skin use. It can be further processed into shampoos, conditioners, and other topical cosmetics.

[0035] This invention first provides the use of the ring (Trp-Arg) in the preparation of cosmetics that promote hair growth.

[0036] The present invention further provides cosmetics including rings (Trp-Arg), which can be shampoos, conditioners, hair treatments, hair creams (hair masks), and other cosmetics used for hair growth and nourishment.

[0037] The ring (Trp-Arg) described in this application can be prepared according to the extraction or synthesis processes provided by existing technologies, or it can be purchased directly.

[0038] The cyclic (Trp-Arg) is usually prepared with excipients to form topical agents, topical medicines, quasi-medicines, or skin cosmetics.

[0039] The additives include bases such as purified water, ethanol, polyols, surfactants, viscosity modifiers, pH adjusters, oily solvents, hair conditioners, vitamins, anti-inflammatory agents, antibacterial agents, cooling agents, moisturizers, antioxidants, metal ion chelating agents, fragrances, etc.

[0040] As a polyol, it can be ethylene glycol, glycerol, 1,3-butanediol, propylene glycol, etc.

[0041] As a surfactant, it can be a nonionic, anionic, or cationic surfactant.

[0042] As viscosity modifiers, cellulose derivatives such as guar gum, hydroxypropyl cellulose, and hydroxypropyl methylcellulose can be selected.

[0043] Citric acid or its salts, tartaric acid or its salts can be used as pH adjusters.

[0044] As an oily solvent, isopropyl palmitate, soybean oil, castor oil, and triglyceride stearate can be selected.

[0045] As a hair quality improver, cyclohexylsiloxanes, etc., can be selected.

[0046] Examples of vitamins include vitamin E, vitamin C, benzyl nicotinate, nicotinamide, D-panthenol, panthenol ether, biotin, pyridoxine hydrochloride, and riboflavin.

[0047] Examples of anti-inflammatory agents include dipotassium glycyrrhizate, β-glycyrrhetinic acid, allantoin, diphenylhydramine hydrochloride, guaiac glycerin ether, and L-menthol.

[0048] Examples of antibacterial and bactericidal agents include glycyrrhizin, ethylparaben, diazolidinyl urea, borax, methylisothiazolinone, trichlorohydroxydiphenyl ether, pyriproxyethanol, triclosan, chlorhexidine gluconate, phenoxyethanol, resorcinol, isopropyl methylphenol, salicylic acid, zinc pyridinethione, benzalkonium chloride, photosensitizer 301, and sodium mononitroguaiacol.

[0049] Examples of cooling agents include L-menthol and camphor.

[0050] Examples of moisturizers include L-pyrrolidone carboxylic acid, sodium hyaluronate, chondroitin sulfate, cordyceps extract, and saffron extract.

[0051] Examples of antioxidants include butylated hydroxyanisole, butylated hydroxytoluene, isopropyl gallate, propyl gallate, and isoascorbic acid.

[0052] Examples of metal ion chelating agents include ethylenediaminetetraacetic acid or its salts.

[0053] As fragrances, examples include natural fragrances such as orange oil, lemon oil, lime oil, and lavender oil, as well as synthetic fragrances such as menthol, rose oxide, linalool, citral, and linalyl acetate.

[0054] Examples of bases for semi-solid or solid dosage forms include petrolatum, solid paraffin, vegetable oil, mineral oil, cetyl alcohol, lanolin, waxes, polyethylene glycol, cholesterol, and white oil. Additives such as lecithin and other emulsifiers, as well as lower alcohols such as ethanol and isopropanol, can be added as needed.

[0055] The beneficial effects of this invention:

[0056] The Trp-Arg ring described in this application, as well as cosmetics containing Trp-Arg, exhibit good hair growth promoting effects when applied to the skin (especially the scalp). Compared with the control group, its hair growth effect increases with increasing concentration. When the concentration reaches 2%, its effect on promoting the proliferation of human outer root sheath cells reaches approximately 111.8%. Its effect on promoting the proliferation of human dermal papilla cells and human epidermal keratinocytes reaches 112.4% and 146.6%, respectively. This application further prepared shampoos, conditioners, and hair creams containing Trp-Arg, and further demonstrated through animal model experiments that cosmetics containing Trp-Arg can effectively promote hair growth, especially scalp hair. Attached Figure Description

[0057] Appendix Figure 1 Hair regrowth effect of different concentrations of Trp-Arg (n=10)

[0058] Appendix Figure 2 Examples 6-8: Hair regrowth effects of various formulas (n=10) Detailed Implementation

[0059] The present invention will be further illustrated below with reference to specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in this technical field.

[0060] Example 1: Skin irritation test on experimental animals after repeated application (conducted according to Chinese Patent CN102526160B).

[0061] I. Materials and Methods

[0062] 1. The test substance is a 10% aqueous solution of cyclohexane (Trp-Arg).

[0063] 2. Test animals: 4 ordinary white New Zealand rabbits

[0064] 3. Experimental Method: 24 hours prior to the experiment, the hair on both sides of the spine on the animal's back was shaved, with a shaving area of ​​approximately 3cm × 3cm on each side. During the experiment, 0.5mL of the test substance was evenly applied to a 2.5cm × 2.5cm area of ​​skin on the left side, with distilled water used as a control on the right side. Application was performed once daily for 7 consecutive days. Starting from the second day, the hair was removed before each application, and the skin was washed with warm water. Skin reactions were observed and scored 1 hour later. After the experiment, the total score over 14 days, the average score for each animal over 14 days, and the average score for each animal each day were calculated, and the intensity of skin irritation was graded.

[0065] II. Scoring Criteria

[0066]

[0067] Skin irritation intensity grading

[0068] Integral mean Intensity 0-0.5 Non-irritating 0.5-2.0 Mildly irritating 2.0-6.0 Moderately irritating 6.0-8.0 Strongly irritating .

[0069] III. Test Results

[0070] Table 1 Results of multiple skin irritation tests

[0071]

[0072] The rating data in the table above shows that the 10% Trp-Arg aqueous solution is safe and non-irritating, and has good safety for topical use on the skin.

[0073] Example 2: Allergic Reaction Test (Referring to the test method in Chinese Patent CN 102429846 B and the 2015 edition of the Cosmetic Safety Technical Specifications)

[0074] I. Materials and Methods

[0075] 1. Preparation method of test substance: 10% Trp-Arg aqueous solution

[0076] 2. Test animals: Common-grade British albino guinea pigs, divided into three groups: test substance experimental group and positive substance experimental group, with 20 animals in each group, and test substance control group, with 20 animals in each group.

[0077] 3. Test methods:

[0078] Induction contact: Apply 0.2 ml of the test substance to the shaved 2 cm × 2 cm area of ​​skin on the left side of the animal in the test substance experimental group, cover with two layers of gauze and one layer of cellophane, fix with adhesive tape, and seal for 6 hours. Repeat the same method on day 7 and day 14. The positive control group was treated with 4.0% 2,4-dinitrochlorophenylacetone solution, and the test substance control group was treated with the same solvent (water).

[0079] Challenge contact: 14 days after the last induction, 0.2 ml of the challenge agent was applied to a 2cm × 2cm area of ​​skin on the right side of both the test agent experimental group and the test agent control group (the area had been shaved 24 hours prior). The area was covered with two layers of gauze and one layer of cellophane, and then sealed with adhesive tape for 6 hours. The positive control group underwent the same challenge contact with a 1.2% 2,4-dinitrochlorophenylacetone solution, while the test agent control group underwent the same treatment with the same solvent. Skin reactions were observed at 24 and 48 hours after the challenge contact, and skin reaction scores and sensitization intensity were determined.

[0080] 4. Experimental Results: The skin reactions of each group of animals at 24 and 48 hours are shown in the table.

[0081] Table 2 Results of the local skin allergy test in guinea pigs

[0082]

[0083] As can be seen from the data in the table above, the 10% Trp-Arg aqueous solution did not induce any allergic reactions in the skin of guinea pigs, indicating good safety for topical application.

[0084] Example 3: Human outer root sheath cell proliferation assay (conducted according to the reference: Zhou Naihui, Fan Weixin, Effect of angiopoietin on the proliferation of human hair follicle outer root sheath keratinocytes, Journal of Clinical Dermatology, 2011, Vol. 40, No. 6, pp. 328-331)

[0085] I. Isolation and culture of keratinocytes from the outer root sheath of human hair follicles:

[0086] Fresh occipital scalp was collected and routinely disinfected. Subcutaneous tissue was extracted and digested overnight at 4°C with 0.5% dissociative enzyme. After washing with PBS (phosphate-buffered saline), the hair follicle epithelium (including the hair shaft, inner root sheath, and outer root sheath) was excised using ophthalmic curved forceps. The excised follicle epithelium was then digested with trypsin cell digestion solution at 37°C for 30 minutes until the outer root sheath was freed into single cells. Digestion was terminated with 0.1% soybean trypsin inhibitor. The mixture was filtered through a 200-mesh sieve, and the single-cell suspension was collected. The suspension was centrifuged at 1000 rpm for 5 minutes. The cell pellet was thoroughly mixed with serum-free keratinocyte culture medium at a concentration of 1.2 × 10⁻⁶. 9 Inoculate at a density of 1 / L at 25cm 2 Cell culture flasks. Change the medium on the third day to remove adherent cells and impurities.

[0087] II. Effects of the Trp-Arg ring on the proliferation of keratinocytes in the outer root sheath of human hair follicles:

[0088] Second-generation human hair follicle outer root sheath keratinocytes in the logarithmic growth phase were digested with 0.25% trypsin and then subjected to 2.0 × 10⁻⁶ ppm. 8 Cells were seeded at a density of 100 μL / L in 96-well plates. After cell attachment, the medium was changed, and different concentrations of Trp-Arg aqueous solution were added. Each concentration was replicated in 6 wells, with 100 μL per well. After culturing for 48 h, cell proliferation was detected by MTT assay.

[0089] MTT assay: Add 15 μL of MTT (5 g / L) to each well of a 96-well plate, incubate at 37°C for 4 hours, then discard the culture medium. Add 150 μL of dimethyl sulfoxide to each well, protect from light, and vortex thoroughly to mix. Measure the absorbance at 490 nm (A490) using a microplate reader, and calculate the cell proliferation percentage using the following formula:

[0090] Cell proliferation percentage (%) = (experimental sample A490 / blank control sample A490-1) * 100%.

[0091] Table 3: Effects of different concentrations of Trp-Arg on the proliferation of human outer root sheath cells (n=6)

[0092] Cyclic (Trp-Arg) concentration A490 mean Cell proliferation percentage (%) Blank control 0.389 / 0.1％ 0.402 3.3％ 0.5％ 0.512 31.6％ 1.0％ 0.765 96.7％ 2.0％ 0.824 111.8％ .

[0093] in conclusion:

[0094] Different concentrations of Trp-Arg were added to human follicular outer root sheath keratinocytes cultured in vitro. After 48 hours of culture, MTT assay showed that the addition of Trp-Arg significantly promoted the proliferation of human follicular outer root sheath keratinocytes. The proliferation activity increased with the increase of the concentration of Trp-Arg added. When 2% Trp-Arg was added, its promoting effect on the proliferation of human outer root sheath cells reached 111.8%.

[0095] Example 4: Hair papilla cell proliferation assay (conducted according to CN 101460138 B)

[0096] (1) Using PromoCell skin follicle dermal papilla cells culture medium, in 96-well plates, at a concentration of 1.0 × 10⁻⁶ 4 Seed hair papilla cells at a cell seeding density of 1 cell / well.

[0097] (2) After culturing at 37°C and 5% carbon dioxide for 1 day, the culture medium was removed, and the cells were washed with DMEM medium containing 1% FCS (fetal bovine serum) for the cell proliferation assay. Then, with deionized water as a blank control, the loop (Trp-Arg) was dissolved in the evaluation medium and added to various concentrations (μmol / L).

[0098] (3) Subsequently, the cells were cultured for 2 days in an environment of 37°C and 5% carbon dioxide gas, and the proliferation rate of hair papilla cells was determined by the MTT method.

[0099] (4) Using the proliferation rate obtained from the medium without added loop (Trp-Arg) as a baseline of 100%, the percentage of cell proliferation was calculated according to the following formula:

[0100] Cell proliferation percentage (%) = (experimental sample A490 / blank control sample A490-1) * 100%.

[0101] The experimental results are shown in the table below:

[0102] Table 4: Effects of different concentrations of Trp-Arg on human dermal papilla cell proliferation (n=6)

[0103] Cyclic (Trp-Arg) concentration A490 mean Cell proliferation percentage (%) Blank control 0.394 / 0.1％ 0.409 3.8％ 0.5％ 0.738 87.3％ 1.0％ 0.761 93.1％ 2.0％ 0.837 112.4％ .

[0104] Example 5: Human epidermal keratinocyte proliferation assay (conducted according to CN 101460138 B)

[0105] (1) EpiLife was used as the basal medium, and 2.0 × 10⁻⁶ mg / L was used in 96-well plates. 4 Cell seeding density of 1 cell / well for seeding human epidermal keratinocytes.

[0106] (2) After culturing at 37°C and 5% carbon dioxide for 1 day, the culture medium was removed, and the cells were washed with MCDB153 medium, an evaluation medium used for cell proliferation assays. Then, using deionized water as a blank control, the loop (Trp-Arg) was dissolved in a similar evaluation medium and added to various concentrations (μmol / L).

[0107] (3) Subsequently, the cells were cultured for 2 days in an environment of 37°C and 5% carbon dioxide gas, and the proliferation rate of human epidermal keratinocytes was determined by the MTT method.

[0108] (4) Using the proliferation rate obtained from the medium without added loop (Trp-Arg) as a baseline of 100%, the percentage of cell proliferation was calculated according to the following formula:

[0109] Cell proliferation percentage (%) = (experimental sample A490 / blank control sample A490-1) * 100%.

[0110] The experimental results are shown in the table below:

[0111] Table 5: Effects of different concentrations of Trp-Arg on the proliferation of human epidermal keratinocytes (n=6)

[0112] Cyclic (Trp-Arg) concentration A490 mean Cell proliferation percentage (%) Blank control 0.365 / 0.1％ 0.416 14.0％ 0.5％ 0.719 97.0％ 1.0％ 0.856 134.5％ 2.0％ 0.900 146.6％ .

[0113] As shown in Examples 4 and 5, different concentrations of Trp-Arg have a significant proliferative effect on dermal papilla cells and human epidermal keratinocytes. The proliferative activity increases with the increase of Trp-Arg concentration. When 2% Trp-Arg is added, its proliferative promotion effect on human dermal papilla cells and human epidermal keratinocytes can reach 112.4% and 146.6%, respectively.

[0114] Example 6: Trp-Arg Shampoo (unit: g)

[0115] Ingredient Formula 1 Formula 2 Formula 3 Formula 4 Behentrimonium chloride 4.0 4.0 4.0 4.0 Guar gum 1.0 1.0 1.0 1.0 Glycol 0.5 0.5 0.5 0.5 Bisimidazolidinyl urea 0.3 0.3 0.3 0.3 Methylisothiazolinone 0.001 0.001 0.001 0.001 Disodium EDTA 3.0 3.0 3.0 3.0 Sodium lauryl sulfate 2.0 2.0 2.0 2.0 Citric acid 1.0 1.0 1.0 1.0 Isopropyl palmitate 3.0 3.0 3.0 3.0 Cyclic (Trp-Arg) 0.1 0.5 1.0 2.0 Deionized water q.s. 100ml 100ml 100ml 100ml .

[0116] Preparation method:

[0117] 1) Take about 70% deionized water, add citric acid, sodium lauryl sulfate, disodium EDTA, diazolidinyl urea, methylisothiazolinone, docosyltrimethylammonium chloride, and cyclohexane (Trp-Arg) in sequence, stir to dissolve, and set aside as an aqueous phase;

[0118] 2) Take isopropyl palmitate and glycyrrhizin, mix them evenly to form the oil phase, and set aside.

[0119] 3) Under stirring conditions, add the aqueous phase to the oil phase, stir, and mix evenly;

[0120] 4) Add the prescribed amount of guar gum to the material obtained in step 3), and bring the volume to 100 ml with deionized water;

[0121] 5) Deaeration and filling produce Trp-Arg shampoo.

[0122] Example 7: Trp-Arg Conditioner (unit: g)

[0123] Ingredient Formula 1 Formula 2 Formula 3 Formula 4 Glycerin 3.0 3.0 3.0 3.0 Vitamin E 0.5 0.5 0.5 0.5 Lauryltrimonium chloride 0.5 0.5 0.5 0.5 Cetyl alcohol 1.0 1.0 1.0 1.0 Hydroxypropyl cellulose 0.5 0.5 0.5 0.5 D-Panthenol 0.1 0.1 0.1 0.1 Cyclohexasiloxane 1.0 1.0 1.0 1.0 Nipagin ethyl ester 0.1 0.1 0.1 0.1 Methylisothiazolinone 0.001 0.001 0.001 0.001 Stearyl glycerin esters 0.8 0.8 0.8 0.8 Isoceteth-20 1.0 1.0 1.0 1.0 Cyclic (Trp-Arg) 0.1 0.5 1.0 2.0 Deionized water q.s. 100ml 100ml 100ml 100ml .

[0124] Preparation method:

[0125] (1) Weigh the raw materials according to the formula.

[0126] (2) Heat 70ml of deionized water, glycerol, D-panthenol, hydroxypropyl cellulose, lauryl trimethylammonium chloride and cyclo(Trp-Arg) in an aqueous phase pot to 60℃, stir well, and then transfer to a vacuum emulsification pot.

[0127] (3) Add cetyl alcohol, cyclohexylsiloxane, ethylparaben, triglyceride stearate and isocetyl alcohol polyether-20 to the oil phase pot and heat to 60°C. Stir evenly and then transfer to a vacuum emulsification pot.

[0128] (4) Start the vacuum device. Homogenize under vacuum for 5 to 10 minutes, then stir and cool.

[0129] (5) When the vacuum emulsifying pot is cooled to 40°C, add vitamin E and methylisothiazolinone, make up the volume with deionized water, and stir evenly.

[0130] (6) Continue cooling to room temperature and discharge the material.

[0131] Example 8: Trp-Arg hair growth formula (unit: g)

[0132] Ingredient Formula 1 Formula 2 Formula 3 Formula 4 Resorcinol 0.3 0.3 0.3 0.3 Cholesterol 1.5 1.5 1.5 1.5 Lecithin 0.5 0.5 0.5 0.5 Vitamin E 0.5 0.5 0.5 0.5 Castor oil 33.0 33.0 33.0 33.0 Borax 2.0 2.0 2.0 2.0 Salicylic acid 0.4 0.4 0.4 0.4 Vitamin C 0.1 0.1 0.1 0.1 White oil 45.5 45.5 45.5 45.5 95% Ethanol 6ml 6ml 6ml 6ml Cyclic (Trp-Arg) 0.1 0.5 1.0 2.0 Deionized water q.s. 100ml 100ml 100ml 100ml .

[0133] Preparation method:

[0134] (1) Crush borax, Trp-Arg, and vitamin C;

[0135] (2) Add the material obtained in step 1) to an appropriate amount of deionized water, stir, heat to 70°C, stir for 30 minutes, cool to 60°C, and set aside.

[0136] (3) Heat 95% ethanol to 60°C and add it to the material obtained in step 2). Keep the temperature constant at 60°C and continue stirring for 30 minutes to form an aqueous phase.

[0137] (4) Heat the white oil to 110°C, maintain for 20 minutes, cool down to 60°C, and add castor oil, cholesterol, lecithin, vitamin E, salicylic acid and resorcinol in sequence. Keep the temperature constant at 60°C to prepare the oil phase for later use.

[0138] (5) Take the aqueous phase obtained in step 3), add it to the oil phase obtained in step 4) under constant temperature of 60℃ and stirring, make up the volume with deionized water, continue stirring and cooling to room temperature, and then release and package the material.

[0139] Example 9: Animal Hair Growth Promotion Test (implemented according to Chinese Patent CN 104254317 A)

[0140] Hair growth promotion experiments were conducted in mice using different concentrations of Trp-Arg and the formulations from Examples 6-8.

[0141] (1) Test sample

[0142] Negative control: Deionized water

[0143] Test samples: different concentrations of Trp-Arg and various formulations from Examples 6-8

[0144] (2) Experimental animals and breeding conditions

[0145] Male C3H mice (18.8g-23.6g, 47-48 weeks old) were used in the experiment. Three days before grouping, the back (excluding the flanks) of the mice from the neck to the tail was shaved with foam shaving cream using a razor.

[0146] The animals were raised in an animal husbandry room with a temperature of 23°C, humidity of 55%, 12 hours of light and darkness, and 12 air changes per hour. They were allowed free access to solid complete feed and tap water.

[0147] (3) Sample application

[0148] Samples were administered percutaneously. Different test samples (controls or test samples) were administered to the dorsal skin of mice using microtubes, gently applied with a finger (10 mice per sample group). The dosage was approximately 100 μl per site.

[0149] The treatment was administered once daily. The first day of administration was designated as day 1, and treatment continued for 24 consecutive days. Two hours after administration, venous blood was collected to measure blood glucose levels, in order to detect the effect of the test sample on blood glucose in the experimental animals.

[0150] (4)Judgment

[0151] For visual observation of the shaved area, scores were given based on the judgment criteria shown in the table below on the grouping days (application day 1), application days 5, 10, 15, 20 and 25 (1 day after all application was completed), and the mean ± standard deviation was calculated for each group.

[0152] Table 6: Judgment Criteria

[0153] Symptom Score Skin appears pink 1 Skin discoloration to gray (less than 30% of shaved area) 2 Skin discoloration to gray (more than 60% of shaved area) or hair elongation observed (less than 30% of shaved area) 3 Skin discoloration to gray (more than 60% of shaved area) or hair elongation observed (less than 60% of shaved area) 4 Hair elongation observed (more than 60% of shaved area) 5 .

[0154] (5) Results

[0155] The results of the hair growth status score observation are shown in Table 7. Figure 1 , Figure 2 The values ​​in the table represent the mean ± standard deviation.

[0156] Table 7. Hair regrowth effects of different concentrations of Trp-Arg and various formulations in Examples 6-8 (n=10)

[0157]

[0158] As shown in Table 7 and Figure 1 , Figure 2 As shown, compared with the negative control group, after 24 days of transdermal application of different concentrations of Trp-Arg and each formulation in Examples 6-8, the hair regrowth status scores of the application groups were significantly higher after 15 days of application, and the difference increased significantly with the extension of application time. It can be confirmed that different concentrations of Trp-Arg and each formulation in Examples 6-8 have different degrees of hair regrowth effects.

[0159] Chinese patent CN106456633A indicates that cyclic tryptophan arginine (compound number 189) possesses GLP-1 agonist activity and can regulate human glucose metabolism. However, it only describes the GLP-1 agonist activity of cyclic (Trp-Arg) through cellular-level experiments, without specifying the exact route of administration or its impact on the GLP-1 agonist activity of cyclic (Trp-Arg). In this experiment, fasting blood glucose levels in mice were monitored 20 days after transdermal administration of different concentrations of cyclic (Trp-Arg) and different formulations of cosmetics from Examples 6-8 to further illustrate the effect of transdermal administration of cyclic (Trp-Arg) on ​​blood glucose levels in experimental animals. The experimental results are shown in Table 8 (n=10).

[0160] As shown in Table 8, statistical analysis revealed that transdermal administration of different concentrations of Trp-Arg aqueous solution and the various formulations of cosmetics from Examples 6-8 did not significantly affect blood glucose levels in mice after 20 days compared to the negative control (deionized water). This indicates that transdermal administration of 0.1-2.0% Trp-Arg and the various formulations of cosmetics from Examples 6-8 had no adverse effects on blood glucose levels in experimental mice.

[0161] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

[0162]

Claims

1. The use of Trp-Arg cyclohexane (TRP-Arg) in the preparation of cosmetics that promote hair growth, characterized in that, The chemical structure of the Trp-Arg ring is as follows:

2. The use as described in claim 1, characterized in that, The mass percentage concentration of the Trp-Arg ring in the cosmetic product is 0.1%-2.0%.

3. The use as described in claim 1, characterized in that, The cosmetic contains 0.5% by mass of the Trp-Arg ring.

4. The use as described in claim 1, characterized in that, The cosmetic contains 1% (by mass) of the Trp-Arg ring.

5. The use as described in claim 1, characterized in that, The cosmetic contains 2.0% by mass of the Trp-Arg ring.

6. The use as described in claim 1, characterized in that, The cosmetics mentioned are shampoos, conditioners, hair treatments, hair lotions, or other cosmetics used for hair growth and nourishment.

7. The use as described in claim 6, characterized in that, The shampoo formula is as follows: 4.0g of dodecyltrimethylammonium chloride Guar gum 1.0g Ganbaosu 0.5g Diazolidinyl urea 0.3g Methylisothiazolinone 0.001g Disodium ethylenediaminetetraacetate 3.0g Sodium lauryl sulfate 2.0g Citric acid 1.0g Isopropyl palmitate 3.0g Cyclic (Trp-Arg) 0.1g-2.0g Bring the volume to 100 mL with deionized water.

8. The use as described in claim 6, characterized in that, The conditioner formula is as follows: 3.0g glycerin Vitamin E 0.5g 0.5g lauryltrimethylammonium chloride Cetyl alcohol 1.0g Hydroxypropyl cellulose 0.5g D-Panthenol 0.1g Cyclohexylsiloxane 1.0g Ethylparaben 0.1g Methylisothiazolinone 0.001g Triglycerides stearate 0.8g Isoceridyl ether-20 1.0g Cyclic (Trp-Arg) 0.1g-2.0g Add deionized water to a final volume of 100 mL.

9. The use as described in claim 6, characterized in that, The formula for the hair cream is as follows: resorcinol 0.3g Cholesterol 1.5g Lecithin 0.5g Vitamin E 0.5g Castor oil 33.0g Borax 2.0g Salicylic acid 0.4g Vitamin C 0.1g 45.5g of white oil 6 mL of 95% ethanol Add 0.1g-2.0g of Trp-Arg to 100mL of deionized water.

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