Anti-hair-loss hair-growing composition, liposome, and preparation method and application of anti-hair-loss hair-growing composition and liposome

Abstract

The invention relates to the technical field of daily chemical products, in particular to an anti-hair-loss hair-nourishing composition, a lipidosome and a preparation method and application of the lipidosome. Aiming at different target spots of androgen-derived alopecia, active matters with different anti-alopecia mechanisms are reasonably matched to obtain the anti-alopecia hair-growing composition, and the composition is prepared into the nano-liposome, so that the flexible application of the composition in dosage forms can be improved, the adsorption time of the preparation on scalp, the deep penetration time of the preparation into hair follicles and the retention time of the preparation in the hair follicles are prolonged, and the hair-growing effect is improved. The bioavailability is improved, and the dual effects of preventing hair loss, growing hair and fixing hair are achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of daily chemical products, and in particular to a hair loss prevention and growth-promoting composition, a liposome, and a preparation method and application thereof. Background Art

[0002] Currently, hair loss is becoming younger, and people's demand for anti-hair loss and hair growth products is increasing day by day. The anti-hair loss market has broad prospects. Androgenic alopecia is the most common cause of hair loss clinically. With the development of science and technology, studies have found that some active substances have good effects in treating hair loss, promoting hair growth and gray hair. For example, moloniside can upregulate Wnt10b, β-catenin and lef1, and play a role in inducing the periodic regeneration of hair follicles. Diaminopyrimidine oxide (Kopexil) and pyrrolidine diaminopyrimidine oxide (Kopyrrol) are newly developed anti-hair loss and hair growth active substances. They have been approved for use in personal care products in Europe, the United States and Japan. Diaminopyrimidine oxide and pyrrolidine diaminopyrimidine oxide are crystalline powders with poor water solubility. The two active ingredients are generally compounded for use in hair growth liquids or shampoo products. The products contain a large amount of alcohols and surfactants, which can easily lead to uncomfortable experiences such as dry scalp and itching after use; secondly, due to the skin's own barrier, the active substances are difficult to penetrate into the hair follicles and cannot directly act on the target site, so the bioavailability is low.

[0003] It is difficult to add them directly to cosmetic preparations. In addition, some polypeptide active ingredients with hair loss prevention and growth effects are sensitive to enzymes, light, heat, pH, etc. If they are added directly into the formula without treatment, they can easily lead to the inactivation of the active ingredients and the instability of the formula.

[0004] Therefore, many anti-hair loss and hair growth products on the market have a single therapeutic target, low bioavailability, and many active ingredients are unstable and easily degraded and inactivated. They also have poor transdermal absorption and are difficult to penetrate into hair follicles, greatly reducing the effectiveness of anti-hair loss and hair growth. Summary of the Invention

[0005] The purpose of the present invention is to overcome the shortcomings of the existing technology and provide an anti-hair loss and hair growth composition prepared into nanoliposomes to achieve the dual effects of preventing hair loss and promoting hair growth and strengthening hair.

[0006] To achieve the above object, the technical solution adopted by the present invention is:

[0007] In a first aspect, the present invention provides an anti-hair loss and growth composition, comprising the following components in parts by weight: 3-6 parts of diaminopyrimidine oxide, 1-5 parts of pyrrolidino diaminopyrimidine oxide, 0.1-1 part of panthenol, 0.1-0.5 parts of adenosine, 0.01-0.1 parts of acetyl tetrapeptide-3, and 0.01-0.5 parts of Cornus officinalis extract.

[0008] The present invention comprehensively considers a variety of anti-hair loss and hair growth mechanisms, and for different targets of androgenic alopecia, scientifically combines diaminopyrimidine oxide, pyrrolidinopyrimidine oxide, panthenol, adenosine, acetyl tetrapeptide-3, and cornus extract to obtain an anti-hair loss and hair growth composition. Diaminopyrimidine oxide (Kopexil) and pyrrolidinopyrimidine oxide (Kopyrrol) are new hair growth promoters, similar in structure to minoxidil, and are approved raw materials for cosmetics. They are safe and have no toxic side effects. The two have different working principles. Kopexil can interfere with the production of mature collagen by fibroblasts and prevent the deposition of mature collagen, thereby avoiding the hardening and shrinkage of the surrounding hair follicles; Kopyrrol can promote the proliferation of hair papilla cells. The synergistic effect of the two active ingredients, diaminopyrimidine oxide and pyrrolidinopyrimidine oxide, can effectively treat androgenic alopecia by stimulating the regeneration of hair follicles, optimizing the hair growth cycle, and accelerating the growth of new hair. Adenosine increases the expression of fibroblast growth factor in dermal papilla cells, thereby dilating blood vessels, increasing blood circulation, and reducing hair loss. Acetyl tetrapeptide-3, a hair loss prevention and hair growth peptide, accelerates the synthesis of extracellular matrix proteins (such as laminin and collagen III), increases the volume and length of hair follicles, strengthens hair roots, repairs the epidermal-dermal connective tissue, and anchors hair within the follicle. Cornus officinalis extract contains large amounts of cornus officinalis and morroniside, which have anti-inflammatory and antioxidant effects and promote microcirculation in hair follicles. They can improve scalp blood supply, inhibit DHT (dihydrotestosterone) damage to hair follicles, and significantly enhance the proliferation and in vitro migration of outer root sheath cells (ORSCs). The combination of these six active ingredients has a balanced effect in preventing hair loss, moisturizing, and repairing hair follicles, synergizing to achieve significant hair loss prevention and hair growth results.

[0009] Preferably, the following components are included in parts by weight: 4-6 parts of diaminopyrimidine oxide, 1.5-5 parts of pyrrolidinopyrimidine oxide, 0.5-1 part of panthenol, 0.2-0.5 part of adenosine, 0.03-0.1 part of acetyl tetrapeptide-3, and 0.05-0.5 part of cornus officinalis extract.

[0010] In a second aspect, the present invention provides an anti-hair loss and hair growth liposome, which comprises the following components in percentage by mass: 4.22-13.1% of the anti-hair loss and hair growth composition and 86.9-95.78% of a carrier excipient.

[0011] Nanoliposomes can encapsulate water-soluble or fat-soluble active ingredients and have the advantages of sustained release, biodegradability, non-immunogenicity and toxicity. Their nanoscale size and good deformability give them a strong ability to penetrate physiological tissue barriers. Nanoliposomes are used in hair care products or transdermal formulations to significantly improve the skin penetration ability of active ingredients. By simultaneously encapsulating effective ingredients with different mechanisms in the same nanoliposome and delivering them through the skin, sustained release and long-term effects can be achieved, while also realizing the synergistic effect of multi-effect and multi-target ingredients, significantly improving efficacy. The present invention prepares the anti-hair loss and hair growth composition into nanoliposomes, which can improve its flexible application in dosage forms, increase the adsorption of the preparation on the scalp and its deep penetration into hair follicles and the time it stays in the hair follicles, and improve bioavailability. The anti-hair loss and hair growth liposomes can enrich the anti-hair loss and hair growth composition in the target area for hair loss prevention and hair growth, allowing the active ingredients to penetrate into the hair follicles, achieving the dual effects of hair loss prevention and hair growth, and hair strengthening.

[0012] Preferably, the carrier excipients include at least one of phospholipids, cholesterol, emulsifiers, polyols, and water.

[0013] Preferably, the phospholipid comprises at least one of soybean lecithin, hydrogenated lecithin, egg yolk lecithin, hydrogenated soybean lecithin, and hydrogenated egg yolk lecithin.

[0014] Preferably, the emulsifier includes at least one of polyoxyethylene castor oil emulsifiers, polyoxyethylene hydrogenated castor oil emulsifiers, polyglycerol emulsifiers, poloxamer, coconut glucoside, triglycerides, pyrrolidones, polyglyceryl esters, polyglyceryl lactate, polyglyceryl stearate, polyglyceryl caprylate / caprate, polyglyceryl laurate, Tween 80, Tween 20, Tween 60, PEG-20 hydrogenated castor oil, PEG-40 hydrogenated castor oil, and PEG-60 hydrogenated castor oil.

[0015] Preferably, the polyol includes at least one of glycerol, propylene glycol, 1,3-butylene glycol, 1,3-propylene glycol, 1,2-pentanediol, ethoxydiglycol, 1,2-hexanediol, dipropylene glycol, isopropyl alcohol, polyethylene glycol-200, PPG-10 sorbitol, octyldodecanol, and ethanol.

[0016] Preferably, the carrier excipients include the following components in parts by weight: 0.5-5 parts of phospholipids, 0.1-1 parts of cholesterol, 2-20 parts of emulsifiers, 10-50 parts of polyols, and 10.9-83.18 parts of water.

[0017] In a third aspect, the present invention provides a method for preparing the anti-hair loss and hair growth liposome, comprising the following steps:

[0018] (1) mixing diaminopyrimidine oxide, pyrrolidino diaminopyrimidine oxide, adenosine, panthenol, cornus officinalis extract, phospholipid, cholesterol, emulsifier and polyol to obtain a mixed solution A;

[0019] (2) mixing acetyl tetrapeptide-3 with water to obtain a mixed solution B;

[0020] (3) Pour mixed solution A into mixed solution B and mix well to obtain mixed solution C;

[0021] (4) Nano-processing the mixed solution C to obtain the anti-hair loss and hair growth liposome.

[0022] Preferably, the mixing condition in step (1) is: stirring evenly at 40-65°C.

[0023] Preferably, the mixing condition in step (2) is: stirring evenly at 45-65°C.

[0024] Preferably, the conditions for uniform mixing in step (3) are: uniform stirring at 45-65°C.

[0025] Preferably, the nano-processing comprises extrusion, ultrasound or high-pressure homogenization.

[0026] Preferably, the high-pressure homogenization condition is: 500-1000 bar.

[0027] In a fourth aspect, the present invention provides the anti-hair loss and hair growth liposome or the use of the anti-hair loss and hair growth liposome in the preparation of an anti-hair loss and hair growth product.

[0028] In a fifth aspect, the present invention provides an anti-hair loss and hair growth product, comprising the anti-hair loss and hair growth liposome or the anti-hair loss and hair growth liposome.

[0029] The beneficial effects of the present invention are:

[0030] The present invention comprehensively considers multiple mechanisms of preventing hair loss and promoting hair growth, and targets different targets of androgenic alopecia. It scientifically combines diaminopyrimidine oxide, pyrrolidinopyrimidine oxide, panthenol, adenosine, acetyl tetrapeptide-3, and Cornus officinalis extract to obtain an anti-hair loss and promoting hair growth composition. The combination of the six active ingredients has a balanced effect in preventing hair loss, moisturizing and repairing hair follicles, and synergistically enhances their effectiveness, thereby achieving significant anti-hair loss and promoting hair growth effects.

[0031] The present invention incorporates nanocarrier technology, employing nanocarriers to encapsulate and deliver the anti-hair loss and hair growth composition. Based on the structure and properties of the nanocarriers, the resulting liposomes exhibit excellent stability and water dispersibility, increasing the solubility of the anti-hair loss and hair growth composition, allowing it to reach a sufficient concentration in the product to achieve its desired effects. This effectively improves the solubility of the ingredients in the anti-hair loss and hair growth composition while providing a stable storage space for them, preventing unnecessary degradation or inactivation of the composition during storage and before use, thereby increasing the concentration of the active ingredients in the anti-hair loss and hair growth product. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 In vitro skin penetration results. DETAILED DESCRIPTION

[0033] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0034] Unless otherwise specified, the experimental methods used in the present invention are all conventional methods, and the materials, reagents, etc. used are commercially available products and can be obtained from commercial channels unless otherwise specified.

[0035] The Cornus officinalis extract used in the present invention was purchased from Xi'an Shilin Technology Co., Ltd. The present invention found in experimental research that the extraction method does not affect its efficacy.

[0036] Example 1:

[0037] An embodiment of the anti-hair loss and hair growth liposome of the present invention; the anti-hair loss and hair growth liposome comprises the following components in percentage by weight:

[0038] Diaminopyrimidine oxide 6%, pyrrolidino diaminopyrimidine oxide 5%, panthenol 1%, adenosine 0.5%, acetyl tetrapeptide-3 0.1%, cornus officinalis extract 0.5%, soy lecithin 5%, cholesterol 1%, cocoyl glucoside 10%, polyglyceryl lactate 10%, 1,2-pentylene glycol 20%, glycerin 20%, propylene glycol 10%, water 10.9%.

[0039] The preparation method of the anti-hair loss and hair growth liposome comprises the following steps:

[0040] (1) diaminopyrimidine oxide, pyrrolidinopyrimidine oxide, adenosine, panthenol, cornus officinalis extract, soybean lecithin, 1,2-pentanediol, glycerol, propylene glycol, cocoyl glucoside, and polyglycerol lactate were mixed and stirred at 65° C. to form a homogeneous and clear mixed solution A;

[0041] (2) Stirring acetyl tetrapeptide-3 and water at 45°C to form a homogeneous and clear mixed solution B;

[0042] (3) Pour mixed solution A into mixed solution B and mix evenly, stirring at 45°C to form a uniform mixed solution C;

[0043] (4) The mixed solution C is subjected to high-pressure homogenization treatment at 1000 bar, the cycle is repeated 3 times, and the mixture is cooled to room temperature to obtain the anti-hair loss and hair growth liposome.

[0044] Example 2:

[0045] An embodiment of the anti-hair loss and hair growth liposome of the present invention; the anti-hair loss and hair growth liposome comprises the following components in percentage by weight:

[0046] Diaminopyrimidine oxide 5%, pyrrolidino diaminopyrimidine oxide 3%, panthenol 0.8%, adenosine 0.3%, acetyl tetrapeptide-3 0.06%, cornus officinalis extract 0.3%, hydrogenated lecithin 3%, cholesterol 0.5%, polysorbate 80 10%, PEG-20 hydrogenated castor oil 5%, 1,2-hexanediol 10%, octyldodecanol 10%, ethoxydiglycol 10%, water 42.04%.

[0047] The preparation method of the anti-hair loss and hair growth liposome comprises the following steps:

[0048] (1) Diaminopyrimidine oxide, pyrrolidinopyrimidine oxide, adenosine, panthenol, cornus officinalis extract, hydrogenated lecithin, 1,2-hexanediol, octyldodecanol, ethoxydiglycol, Tween 80, and PEG-20 hydrogenated castor oil were mixed and stirred at 65° C. to form a homogeneous and clear mixed solution A;

[0049] (2) Stirring acetyl tetrapeptide-3 and water at 45°C to form a homogeneous and clear mixed solution B;

[0050] (3) Pour mixed solution A into mixed solution B and mix evenly, stirring at 45°C to form a uniform mixed solution C;

[0051] (4) The mixed solution C is subjected to high-pressure homogenization treatment at 800 bar, the cycle is repeated 3 times, and the mixture is cooled to room temperature to obtain the anti-hair loss and hair growth liposome.

[0052] Example 3

[0053] An embodiment of the anti-hair loss and hair growth liposome of the present invention; the anti-hair loss and hair growth liposome comprises the following components in percentage by weight:

[0054] Diaminopyrimidine oxide 4%, pyrrolidino diaminopyrimidine oxide 1.5%, panthenol 0.5%, adenosine 0.2%, acetyl tetrapeptide-3 0.03%, cornus officinalis extract 0.05%, egg yolk lecithin 1%, cholesterol 0.3%, PEG-20 hydrogenated castor oil 10%, poloxamer 5%, 1,2-pentanediol 10%, butylene glycol 10%, water 57.42%.

[0055] The preparation method of the anti-hair loss and hair growth liposome comprises the following steps:

[0056] (1) Diaminopyrimidine oxide, pyrrolidinopyrimidine oxide, adenosine, panthenol, cornus officinalis extract, egg yolk lecithin, 1,2-pentanediol, butylene glycol, PEG-20 hydrogenated castor oil, and poloxamer were mixed and stirred at 65° C. to form a homogeneous and clear mixed solution A;

[0057] (2) Stirring acetyl tetrapeptide-3 and water at 45°C to form a homogeneous and clear mixed solution B;

[0058] (3) Pour mixed solution A into mixed solution B and mix evenly, stirring at 45°C to form a uniform mixed solution C;

[0059] (4) The mixed solution C is subjected to high-pressure homogenization treatment at 600 bar, the cycle is repeated 3 times, and the mixture is cooled to room temperature to obtain the anti-hair loss and hair growth liposome.

[0060] Example 4

[0061] An embodiment of the anti-hair loss and hair growth liposome of the present invention; the anti-hair loss and hair growth liposome comprises the following components in percentage by weight:

[0062] Diaminopyrimidine oxide 3%, pyrrolidino diaminopyrimidine oxide 1%, panthenol 0.1%, adenosine 0.1%, acetyl tetrapeptide-3 0.01%, cornus officinalis extract 0.01%, hydrogenated soy lecithin 0.5%, cholesterol 0.1%, PEG-20 hydrogenated castor oil 1%, polyglyceryl laurate 1%, 1,2 hexanediol 5%, 1,3 propylene glycol 5%, water 83.18%.

[0063] The preparation method of the anti-hair loss and hair growth liposome comprises the following steps:

[0064] (1) diaminopyrimidine oxide, pyrrolidinopyrimidine oxide, adenosine, panthenol, cornus officinalis extract, hydrogenated soy lecithin, 1,2-hexanediol, 1,3-propylene glycol, PEG-20 hydrogenated castor oil, and polyglyceryl laurate were mixed and stirred at 65° C. to form a homogeneous and clear mixed solution A;

[0065] (2) Stirring acetyl tetrapeptide-3 and water at 45°C to form a homogeneous and clear mixed solution B;

[0066] (3) Pour mixed solution A into mixed solution B and mix evenly, stirring at 65°C to form a uniform mixed solution C;

[0067] (4) The mixed solution C is subjected to high-pressure homogenization treatment at 500 bar, the cycle is repeated 3 times, and the mixture is cooled to room temperature to obtain the anti-hair loss and hair growth liposome.

[0068] Comparative Example 1:

[0069] A comparative example of the anti-hair loss and hair growth liposomes of the present invention; the anti-hair loss and hair growth liposomes of this comparative example differ from those of Example 1 only in that adenosine is not added and is replaced by an equal amount of water, and the remaining components are the same as those of Example 1.

[0070] The preparation method of the anti-hair loss and hair growth liposome is the same as that in Example 1.

[0071] Comparative Example 2:

[0072] A comparative example of the anti-hair loss and hair growth liposome of the present invention; the anti-hair loss and hair growth liposome of this comparative example is different from that of Example 1 only in that panthenol is not added and is replaced by an equal amount of water, and the remaining components are the same as those of Example 1.

[0073] The preparation method of the anti-hair loss and hair growth liposome is the same as that in Example 1.

[0074] Comparative Example 3:

[0075] A comparative example of the anti-hair loss and hair growth liposomes of the present invention; the anti-hair loss and hair growth liposomes of this comparative example differ from those of Example 1 only in that no Cornus officinalis extract is added and an equal amount of water is used instead, and the remaining components are the same as those of Example 1.

[0076] The preparation method of the anti-hair loss and hair growth liposome is the same as that in Example 1.

[0077] Comparative Example 4:

[0078] A comparative example of the anti-hair loss and hair growth liposome of the present invention; the anti-hair loss and hair growth liposome of this comparative example is different from that of Example 1 only in that acetyl tetrapeptide-3 is not added and is replaced by an equal amount of water, and the remaining components are the same as those of Example 1.

[0079] The preparation method of the anti-hair loss and hair growth liposome is the same as that in Example 1.

[0080] Comparative Example 5:

[0081] A comparative example of the anti-hair loss and hair growth liposome of the present invention; the anti-hair loss and hair growth liposome of this comparative example is different from that of Example 1 only in that diaminopyrimidine oxide is not added and is replaced by an equal amount of water, and the remaining components are the same as those of Example 1.

[0082] The preparation method of the anti-hair loss and hair growth liposome is the same as that in Example 1.

[0083] Comparative Example 6:

[0084] A comparative example of the anti-hair loss and hair growth liposome of the present invention; the anti-hair loss and hair growth liposome of this comparative example is different from that of Example 1 only in that pyrrolidinodiaminopyrimidine oxide is not added and is replaced by an equal amount of water, and the remaining components are the same as those of Example 1.

[0085] The preparation method of the anti-hair loss and hair growth liposome is the same as that in Example 1.

[0086] Comparative Example 7:

[0087] A comparative example of the anti-hair loss and hair growth liposome of the present invention; the anti-hair loss and hair growth liposome of this comparative example comprises the following components in percentage by weight:

[0088] Diaminopyrimidine oxide 6.5%, pyrrolidinopyrimidine oxide 5%, panthenol 0.5%, adenosine 0.1%, acetyl tetrapeptide-3 0.1%, cornus officinalis extract 0.9%, and other ingredients are the same as in Example 1 (water is added to 100%).

[0089] The preparation method of the anti-hair loss and hair growth liposome is the same as that in Example 1.

[0090] Comparative Example 8:

[0091] A comparative example of the anti-hair loss and hair growth liposomes of the present invention; the anti-hair loss and hair growth liposomes of this comparative example are different from those of Example 1 only in that the Cornus officinalis extract is replaced with an equal amount of ginseng extract, and the remaining components are the same as those of Example 1.

[0092] The preparation method of the anti-hair loss and hair growth liposome is the same as that in Example 1.

[0093] Comparative Example 9:

[0094] This comparative example provides a free anti-hair loss and hair growth composition; the free anti-hair loss and hair growth composition comprises the following components:

[0095] Diaminopyrimidine oxide 6%, pyrrolidinodiaminopyrimidine oxide 5%, panthenol 1%, adenosine 0.5%, acetyl tetrapeptide-3 0.1%, cornus officinalis extract 0.5%, dimethyl sulfoxide (DMSO) 69.52%, water 17.38%.

[0096] The preparation method of the anti-hair loss and hair growth liposome comprises the following steps:

[0097] The free anti-hair loss and growth composition is obtained by mixing diaminopyrimidine oxide, pyrrolidinopyrimidine oxide, panthenol, adenosine, acetyl tetrapeptide-3, cornus officinalis extract, dimethyl sulfoxide (DMSO) and water.

[0098] Test Example 1: Characterization Test and Stability Test

[0099] Test samples: the anti-hair loss and hair growth liposomes prepared in Examples 1-4 and Comparative Examples 1-8.

[0100] 1. Use an instrument (particle size analyzer) to test the particle size and PDI of the test sample.

[0101] The results showed that the average particle size of the anti-hair loss and hair growth liposomes in Examples 1-4 and Comparative Examples 1-8 was within 10-200 nm, with a PDI of 0.05-0.4. Specifically, the average particle size of the anti-hair loss and hair growth liposomes in Example 1 was 48.98 nm, with a PDI of 0.276; the average particle size of the anti-hair loss and hair growth liposomes in Example 2 was 53.65 nm, with a PDI of 0.295; the average particle size of the anti-hair loss and hair growth liposomes in Example 3 was 52.92 nm, with a PDI of 0.265; and the average particle size of the anti-hair loss and hair growth liposomes in Example 4 was 54.36 nm, with a PDI of 0.272.

[0102] The average particle size of the anti-hair loss and hair growth liposomes in comparative example 1 is 56.89 nm, and the PDI is 0.302; the average particle size of the anti-hair loss and hair growth liposomes in comparative example 2 is 55.98 nm, and the PDI is 0.293; the average particle size of the anti-hair loss and hair growth liposomes in comparative example 3 is 47.12 nm, and the PDI is 0.286; the average particle size of the anti-hair loss and hair growth liposomes in comparative example 4 is 63.31 nm, and the PDI is 0.316; the average particle size of the anti-hair loss and hair growth liposomes in comparative example 5 is 59.65 nm, and the PDI is 0.306; the average particle size of the anti-hair loss and hair growth liposomes in comparative example 6 is 61.62 nm, and the PDI is 0.296; the average particle size of the anti-hair loss and hair growth liposomes in comparative example 7 is 68.82 nm, and the PDI is 0.283; the average particle size of the anti-hair loss and hair growth liposomes in comparative example 8 is 67.15 nm, and the PDI is 0.331.

[0103] 2. Place each test sample in a sealed container at -20°C, room temperature, 4°C, and 45°C for 3 months each, and observe changes in their stability.

[0104] The results showed that the anti-hair loss and hair growth liposomes of Examples 1-4 and Comparative Examples 1-8 did not show stratification or precipitation after being placed in different environments, and the particle size did not change significantly, indicating that the anti-hair loss and hair growth liposomes prepared by the present invention have good stability.

[0105] Test Example 2: Evaluation of irritation to chicken embryo chorioallantoic membrane

[0106] Test sample: the anti-hair loss and hair growth liposomes prepared in Examples 1-4.

[0107] After diluting the test samples 10-fold with water, 200 μL of the dilution solution was dripped onto the surface of the chick embryo chorioallantoic membrane. After waiting for 300 seconds, the changes in the blood vessels were observed. The changes in the CAM blood vessels were observed within 5 minutes, and the initial time of congestion, bleeding, and coagulation in the CAM blood vessels was recorded. The irritation score (IS) was calculated according to the following formula:

[0108] IS=[(301-secH)×5+(301-secL)×7+(301-secC)×9] / 300

[0109] In the above formula, secH is the initial time of congestion, s; secL is the initial time of bleeding, s; secC is the initial time of coagulation, s.

[0110] The mean IS of repeated tests was calculated, and the irritation of the test substances was graded according to the size of the mean, where 0-0.9, 1.0-4.9, 5.0-8.9 and 9-21.0 were classified as no irritation, slight irritation, moderate irritation and severe irritation, respectively.

[0111] The test results showed that after the anti-hair loss and hair growth liposomes diluted 10-fold with water were in contact with the chicken embryo chorioallantoic membrane for 300 seconds, there was no capillary bleeding, vascular melting, or coagulation. The anti-hair loss and hair growth liposomes prepared in Examples 1-4 were tested, and the reaction integrals were 0.06, 0.08, 0.08, and 0.07, respectively, indicating that the anti-hair loss and hair growth liposomes prepared by the present invention are safe and non-irritating.

[0112] Test Example 3: Patch Test

[0113] Test sample: the anti-hair loss and hair growth liposomes prepared in Examples 1-4.

[0114] Thirty subjects were selected and a 10% diluted anti-hair loss and growth liposome solution and a blank control were applied to the flexed side of their forearms for 24 hours. After removing the patch tester, the skin reaction was observed 30 minutes after the indentation disappeared. Skin reactions were also observed 24 and 48 hours after the patch tester was removed.

[0115] The results showed that none of the 30 subjects developed symptoms such as light red spots, erythema, edematous erythema, significant redness and swelling, infiltration or papules, and papules or herpes, indicating that the anti-hair loss and hair growth liposomes prepared by the present invention are non-irritating to human skin.

[0116] Test Example 4: In vitro skin penetration test

[0117] Free RhoB and RhoB nanocarriers: The active ingredient in the anti-desorption liposomes prepared in Example 1 was fluorescently labeled with Rhodamine B (RhoB), and a free Rhodamine B solution of equal concentration was prepared as a control sample (free RhoB).

[0118] A vertical Franz diffusion cell method was used to conduct an in vitro porcine skin permeation experiment. The skin was fixed between the receiving chamber and the donor chamber, and the experiment was divided into two groups, namely the free RhoB group and the RhoB nanocarrier group. 0.5 g each of the RhoB nanocarrier (labeled anti-desorption liposome prepared in Example 1) compound essence (10% RhoB nanocarrier) and the free RhoB compound essence (consistent with the RhoB concentration in the RhoB nanocarrier) were placed in the donor chamber, PBS was used as the receiving solution, and the mixture was stirred and diffused at 37°C. After 2 h and 4 h, the residual sample on the skin was gently wiped off, the skin in the target area was removed, the skin was rinsed again, and the residual moisture was wiped off after thorough cleaning. The sample was frozen and sectioned, and the sections were observed under a laser confocal microscope, and representative areas were selected for photography.

[0119] The results are as follows Figure 1 As shown, free RhoB was primarily concentrated in the stratum corneum at 2 hours and failed to penetrate the stratum corneum barrier. However, the RhoB nanocarriers had already penetrated the stratum corneum barrier and entered the active epidermis by 2 hours. Over time, the fluorescence penetration depth of the RhoB nanocarriers increased, reaching a depth of 264.3 μm at 4 hours, indicating that they had reached the dermis. These results demonstrate that, over the same time period, the fluorescence intensity and penetration depth of the RhoB nanocarriers in the skin were significantly greater than those of free RhoB, demonstrating that the nanocarriers can effectively deliver the encapsulated active ingredients to the deeper layers of the skin, targeting hair follicles.

[0120] Test Example 5 In vitro hair follicle growth test

[0121] Test samples: the anti-hair loss and hair growth liposomes prepared in Examples 1-4 and Comparative Examples 1-8, and the free anti-hair loss and hair growth composition prepared in Comparative Example 9.

[0122] Mouse hair follicles were isolated and cultured in vitro. Starting the next day, the anti-delipidated liposomes were diluted 5000-fold with culture medium. A blank and positive control, minoxidil, were added at a concentration of 10 μM. The cells were incubated in a 37°C, 5% CO2 incubator for 14 days. Vibrissae follicle length was measured and photographed every 7 days. Data are presented as mean ± SD.

[0123] The relative growth rate of hair follicles was calculated according to the following formula:

[0124] Relative growth rate of hair follicles (%) = (average length of hair follicles in each group D14 - average length of hair follicles in each group D0) / (length of hair follicles in the blank control group D14 - length of hair follicles in the blank control group D0) × 100%

[0125] Table 1

[0126]

[0127]

[0128] Note: # Indicates p < 0.01 compared with the blank group; @ indicates p < 0.01 compared with the minoxidil group; a a) Compared with Comparative Examples 1-6, p<0.01; b) Compared with Comparative Example 7, p<0.01; c) Compared with Comparative Example 8, p<0.01; d) Compared with Comparative Example 9, p<0.01.

[0129] The results are shown in Table 1. The hair follicle growth value of the minoxidil group at 14 days was significantly greater than that of the blank control group, indicating that the in vitro culture experiment of mouse hair follicles was successful.

[0130] Compared with the minoxidil group, the anti-hair loss and hair growth liposomes prepared in Examples 1-4 were comparable to or even stronger than minoxidil, and could significantly promote hair follicle growth.

[0131] Compared with the anti-hair loss and hair growth liposomes prepared in Comparative Examples 1-6, the liposomes prepared in Example 1 by compounding the six active ingredients of diaminopyrimidine oxide, pyrrolidinopyrimidine oxide, panthenol, adenosine, acetyl tetrapeptide-3, and Cornus officinalis extract had a better hair follicle growth effect than any combination of the five active ingredients (p<0.01), indicating that the combined use of the active ingredients has a synergistic effect.

[0132] Compared with Comparative Example 7, the weight ratio of aminopyrimidine oxide, pyrrolidinodiaminopyrimidine oxide, panthenol, adenosine, acetyl tetrapeptide-3, and Cornus officinalis extract in the anti-hair loss and hair growth liposomes of Comparative Example 7 is unbalanced, resulting in a decreased effect on hair follicle growth and failure to achieve the optimal effect.

[0133] Compared with Comparative Example 8, Example 1 shows that the anti-hair loss and hair growth liposomes in Comparative Example 8 replace Cornus officinalis extract with ginseng extract, resulting in a decreased hair follicle growth effect. This may be because ginseng extract lacks the DHT antagonism, hair follicle cycle regulation, and synergistic anti-fibrosis effects unique to Cornus officinalis, resulting in a weakened multi-target anti-hair loss mechanism formed by the combination of the six active ingredients. This indicates that replacing any component in the anti-hair loss and hair growth composition of the present invention will affect the effect of hair follicle growth and fail to achieve optimal efficacy. The present invention combines the six specific components of diaminopyrimidine oxide, pyrrolidino diaminopyrimidine oxide, panthenol, adenosine, acetyl tetrapeptide-3, and Cornus officinalis extract to achieve optimal synergistic effects and more significant hair follicle growth efficacy.

[0134] Compared with Comparative Example 9, the hair follicle growth efficacy of the encapsulated nanocomposition of Example 1 is greatly enhanced compared with the free composition.

[0135] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A hair loss prevention and growth composition, characterized in that: The invention comprises the following components in parts by weight: 3-6 parts of diaminopyrimidine oxide, 1-5 parts of pyrrolidino diaminopyrimidine oxide, 0.1-1 part of panthenol, 0.1-0.5 part of adenosine, 0.01-0.1 part of acetyl tetrapeptide-3 and 0.01-0.5 part of cornus officinalis extract.

2. The anti-hair loss and growth-promoting composition according to claim 1, wherein The invention comprises the following components in parts by weight: 5-6 parts of diaminopyrimidine oxide, 3-5 parts of pyrrolidino diaminopyrimidine oxide, 0.8-1 part of panthenol, 0.3-0.5 part of adenosine, 0.06-0.1 part of acetyl tetrapeptide-3 and 0.3-0.5 part of cornus officinalis extract.

3. A hair loss prevention and hair growth liposome, characterized in that: The anti-hair loss and hair growth liposomes include the following components in percentage by mass: 4.22-13.1% of the anti-hair loss and growth composition according to any one of claims 1 to 2 and 86.9-95.78% of carrier excipients.

4. The anti-hair loss and hair growth liposome according to claim 3, wherein: The carrier auxiliary material includes at least one of phospholipid, cholesterol, emulsifier, polyol and water.

5. The anti-hair loss and hair growth liposome according to claim 4, wherein: The phospholipids include at least one of soybean lecithin, hydrogenated lecithin, egg yolk lecithin, hydrogenated soybean lecithin, and hydrogenated egg yolk lecithin; And / or, the emulsifier includes at least one of polyoxyethylene castor oil emulsifiers, polyoxyethylene hydrogenated castor oil emulsifiers, polyglycerol emulsifiers, poloxamer, cocoyl glucoside, triglycerides, pyrrolidones, polyglyceryl esters, polyglyceryl lactate, polyglyceryl stearate, polyglyceryl caprylate / caprate, polyglyceryl laurate, Tween 80, Tween 20, Tween 60, PEG-20 hydrogenated castor oil, PEG-40 hydrogenated castor oil, and PEG-60 hydrogenated castor oil; And / or, the polyol includes at least one of glycerol, propylene glycol, 1,3-butylene glycol, 1,3-propylene glycol, 1,2-pentanediol, ethoxydiglycol, 1,2-hexanediol, dipropylene glycol, isopropyl alcohol, polyethylene glycol-200, PPG-10 sorbitol, octyldodecanol, and ethanol.

6. The anti-hair loss and hair growth liposome according to claim 4, wherein: The carrier auxiliary material comprises the following components in parts by weight: 0.5-5 parts of phospholipid, 0.1-1 parts of cholesterol, 2-20 parts of emulsifier, 10-50 parts of polyol, and 11.9-83.28 parts of water.

7. The method for preparing the anti-hair loss and hair growth liposome according to any one of claims 3 to 6, characterized in that: The steps include: (1) mixing diaminopyrimidine oxide, pyrrolidino diaminopyrimidine oxide, adenosine, panthenol, cornus officinalis extract, phospholipid, cholesterol, emulsifier and polyol to obtain a mixed solution A; (2) mixing acetyl tetrapeptide-3 with water to obtain a mixed solution B; (3) Pour mixed solution A into mixed solution B and mix well to obtain mixed solution C; (4) Nano-processing the mixed solution C to obtain the anti-hair loss and hair growth liposome.

8. The preparation method according to claim 7, wherein The nano-processing includes extrusion, ultrasound or high-pressure homogenization.

9. Use of the anti-hair loss and hair growth composition according to any one of claims 1 to 2 or the anti-hair loss and hair growth liposome according to any one of claims 3 to 6 in the preparation of an anti-hair loss and hair growth product.

10. A hair loss prevention and growth product, characterized in that: The method comprises the anti-hair loss and hair growth composition according to any one of claims 1 to 2 or the anti-hair loss and hair growth liposome according to any one of claims 3 to 6.

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