Exosome compound preparation with hair growth effect and preparation method thereof

Abstract

The invention relates to the technical field of exosomes, in particular to an exosome compound preparation with a hair growing effect and a preparation method of the exosome compound preparation. Comprising 12 to 22 parts of ginsenoside Rb1, 15 to 12 parts of biotin tripeptide-15, 8 to 15 parts of diammonium glycyrrhizinate, 50 to 70 parts of fish collagen peptide, 10 to 20 parts of soybean peptide, 0.5 to 4.5 parts of SHH protein, 0.5 to 2.5 parts of SMO inhibitor, 0.5 to 1.5 parts of HhSP protein, 5 to 10 parts of N-acetyl tyrosine, 0.1 to 2 parts of hyaluronic acid, 0.1 to 1 part of melatonin, 0.1 to 1 part of quercetin, 0.1 to 1 part of kaempferol, 0.1 to 1 part of tanshinone, 0.1 to 1 part of biotin and 0.1 to 1 part of vitamin A. The compound preparation is combined with exosomes from various sources, so that the hair papilla cell function is synergistically enhanced, and the damaged hair follicle structure is repaired.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of hair growth agent preparation, and more particularly to an exosome composite preparation with hair growth efficacy and a preparation method thereof. BACKGROUND

[0002] Hair loss has been a widely concerned topic worldwide, affecting the quality of life of millions or even tens of millions of people. Hair loss not only affects the appearance, but also may cause psychological burden and social adaptation problems to patients. At present, the solutions to hair loss on the market mainly include drug treatment, hair transplantation surgery and various hair growth agents. However, the existing hair growth agents still have many deficiencies in effect and safety.

[0003] At present, exosomes need to be stored in an environment close to the osmotic pressure of the human body, otherwise the phospholipid bilayer structure is easily damaged, resulting in inactivation of internal proteins and miRNA and reducing the curative effect. Although natural exosomes can home to specific tissues, the targeting efficiency is limited. It is necessary to rely on composite carriers (such as hydrogel, microneedle patch) to enhance local permeability, which increases the technical complexity. SUMMARY

[0004] The present application provides an exosome composite preparation with hair growth efficacy and a preparation method thereof. The exosomes activate the Wnt / β-catenin signaling pathway through carrying active molecules, promote the proliferation and differentiation of hair follicle stem cells, accelerate the transition of hair follicles from the resting phase to the growth phase, and delay the involution phase. The composite preparation combines exosomes from multiple sources to synergistically enhance the function of hair papilla cells and repair damaged hair follicle structure.

[0005] In a first aspect, the present application provides an exosome composite preparation with hair growth efficacy, which comprises the following components: 12-22 parts of ginsenoside Rb1, 5-12 parts of biotin tripeptide-1, 8-15 parts of diammonium glycyrrhizinate, 50-70 parts of fish collagen peptide, 10-20 parts of soybean peptide, 12-25 parts of stem cell exosome, 0.5-4.5 parts of SHH protein, 0.5-2.5 parts of SMO inhibitor, 0.5-1.5 parts of HhSP protein, 5-10 parts of N-acetyl tyrosine, 0.1-2 parts of hyaluronic acid, 0.1-1 part of melatonin, 0.1-1 part of quercetin, 0.1-1 part of kaempferol, 0.1-1 part of tanshinone, 0.1-1 part of biotin and 0.1-1 part of vitamin A.

[0006] Preferably, the substance comprises the following components: 18~22 parts of ginsenoside Rb1, 18~12 parts of biotin tripeptide-1, 9~13 parts of diammonium glycyrrhizinate, 65~70 parts of fish collagen peptide, 12~20 parts of soybean peptide, 15~25 parts of stem cell exosome, 0.5~3.5 parts of SHH protein, 0.8~2.5 parts of SMO inhibitor, 5~10 parts of N-acetyl tyrosine, 0.1~2 parts of hyaluronic acid, 0.1~1 part of melatonin, 0.1~1 part of quercetin, 0.1~1 part of kaempferol, 0.1~1 part of tanshinone, 0.1~1 part of biotin and 0.1~1 part of vitamin A.

[0007] Preferably, the preparation method of the biotin tripeptide-1 is as follows: dissolving biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide in tetrahydrofuran, heating to 60~90℃, refluxing for 1~3h to obtain biotin tripeptide-1; the mass ratio of the biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide is 5~8:2~3:1.

[0008] Preferably, the fish collagen peptide is prepared by extracting with protease for 4~9h, adding complex protease after enzyme inactivation for secondary enzymolysis, and the complex protease comprises at least one of bromelain, neutral protease, alkaline protease and flavor protease.

[0009] Preferably, the enzymolysis conditions of the fish collagen peptide are as follows: the temperature is 37~60℃, the pH is 1.5~8.5, the enzymolysis time is 1~12h, and the enzyme addition amount is 0.5%~8% of the raw material.

[0010] Preferably, the stem cell exosome is ipsc-derived, and the total yield of the stem cell exosome is 2.5×10 4 ; the HhSP protein is a hedgehog pathway protein.

[0011] In the second aspect, the present application provides a preparation method of an exosome composite preparation with hair growth effect, comprising the following steps: (1) treating N-acetyl tyrosine, hyaluronic acid, melatonin, quercetin, kaempferol, tanshinone, biotin and vitamin A into stem cell exosome by ultrasonic treatment to obtain exosome-loaded active extract; (2) mixing biotin tripeptide-1, fish collagen peptide, soybean peptide and the exosome-loaded active extract to prepare active peptide-loaded preparation; (3) mixing ginsenoside Rb1, SHH protein, SMO inhibitor, HhSP protein and diammonium glycyrrhizinate with the active peptide-loaded preparation to prepare a mixed composite preparation.

[0012] In a third aspect, the present application provides an application of the exosome composite preparation with hair growth efficacy in the preparation of a product for preventing hair loss, promoting hair growth and hair growth.

[0013] Preferably, the product for preventing hair loss, promoting hair growth and hair growth is a drug or a care product; and / or, The dosage form of the product for preventing hair loss, promoting hair growth and hair growth includes any one of a coating agent, a spray, a lotion, a paste, a rubbing agent and a gel.

[0014] In summary, the present application has the following beneficial effects: 0、The present application regulates the activity and homing of bulge region hair follicle stem cells (HFSCs), ensures the steady state of the hair follicle "growth period (Anagen) - regression period (Catagen) - resting period (Telogen)" cycle, and when the skin is damaged, the pathway can also mobilize HFSCs to migrate to the damaged site and participate in epidermal repair. Exogenous supplement of SHH protein shortens the resting period of hair follicle, and induces the hair follicle in the resting period to enter the growth period in advance; after blocking the HH pathway by using SMO inhibitor, the hair follicle growth will be arrested in the resting period, further confirming the irreplaceability of the HH pathway in the hair growth process. The exosome activates the Wnt / β-catenin signaling pathway through the active molecules carried by it, promotes the proliferation and differentiation of hair follicle stem cells, accelerates the transition of hair follicle from the resting period to the growth period, and delays the regression period. The composite preparation combines exosomes from multiple sources to synergistically enhance the function of hair papilla cells and repair damaged hair follicle structure.

[0015] 2、The present application has natural biocompatibility: derived from lipid bilayer vesicles secreted by cells, no immunogenicity, can avoid local irritation of traditional drugs; efficient targeted delivery: through genetic engineering modification of surface targeting molecules, can accurately recognize hair follicle cells, and deliver multiple proteins to DP and bulge region; stable activity retention: the vesicle structure can protect the protein from enzymatic hydrolysis, and ensure its continuous function in the hair follicle microenvironment.

[0016] 3. This invention achieves hair follicle regeneration at the mechanistic level, exhibiting precision and targeting the core of hair follicle function. Engineered exosomes can penetrate the dermis, directly delivering HhSP protein to DP and HFSCs, activating the HH signaling pathway. This avoids the limitations of traditional hair growth solutions that only have a "surface effect," achieving "awakening hair follicles from the root." It also has a long-lasting effect, extending the hair follicle growth phase. HhSP protein maintains the continuous activation of GLI protein, prolonging the hair follicle growth phase. Clinical data shows that after use, hair density increases by 42%, hair diameter increases by 23%, and the effect lasts for over 12 months. Furthermore, it is safe, with low risk and no side effects. Both exosomes and HhSP protein are natural biomolecules, without the cytotoxicity of chemical drugs. Clinical observations have not revealed side effects such as scalp redness and itching, demonstrating superior biocompatibility compared to traditional hair growth drugs.

[0017] It should be understood that the above general description and the following detailed description are exemplary and explanatory only, and do not limit the scope of protection of the present invention. Detailed Implementation

[0018] The present invention will be further described in detail below with reference to the embodiments. It should be noted that: unless otherwise specified, the conditions in the following embodiments are carried out according to conventional conditions or the conditions recommended by the manufacturer. Unless otherwise specified, the raw materials used in the following embodiments can be obtained from commercially available sources.

[0019] Example Example 1 An exosome compound preparation with hair growth effect comprises the following components: 12 parts ginsenoside Rb1, 5 parts biotin tripeptide-1, 8 parts diammonium glycyrrhizate, 50 parts fish collagen peptide, 10 parts soybean peptide, 12 parts stem cell exosomes, 0.5 parts SHH protein, 0.5 parts SMO inhibitor, 0.5 parts HhSP protein, 5 parts N-acetyltyrosine, 0.1 parts hyaluronic acid, 0.1 parts melatonin, 0.1 parts quercetin, 0.1 parts kaempferol, 0.1 parts tanshinone, 0.1 parts biotin, and 0.1 parts vitamin A.

[0020] The preparation method of biotin tripeptide-1 is as follows: biotin, N-hydroxysuccinimide and 1,3-dicyclohexylcarbodiimide are dissolved in tetrahydrofuran, heated to 60℃ and refluxed for 1 h to obtain biotin tripeptide-1; the mass ratio of biotin, N-hydroxysuccinimide and 1,3-dicyclohexylcarbodiimide is 5:2:1.

[0021] Fish collagen peptides were obtained by extracting with protease for 4 hours, followed by enzyme inactivation and secondary enzymatic hydrolysis with a complex protease, which included bromelain, neutral protease, alkaline protease and flavor protease in a mass ratio of 1:1:1:2.

[0022] The enzymolysis conditions in fish collagen peptide are: the temperature is 37℃, the pH is 1.5, the enzymolysis time is 1h, and the enzyme addition amount is 0.5% of the raw material.

[0023] The stem cell exosome is extracted from the adipose tissue near the rat groin and testis.

[0024] A preparation method of an exosome composite preparation with hair growth efficacy, comprising the following steps: (1) N-acetyl tyrosine, hyaluronic acid, melatonin, quercetin, kaempferol, tanshinone, biotin and vitamin A are treated into stem cell exosomes by ultrasonic treatment to obtain exosome-loaded active extract; (2) Biotin tripeptide-1, fish collagen peptide and soybean peptide are mixed with the exosome-loaded active extract to prepare a loaded active peptide; (3) Ginsenoside Rb1, SHH protein, SMO inhibitor, HhSP protein and diammonium glycyrrhizinate are mixed with the loaded active peptide to prepare a mixed composite preparation.

[0025] Example 2 An exosome composite preparation with hair growth efficacy, comprising the following components: 14 parts of ginsenoside Rb1, 6 parts of biotin tripeptide-1, 9 parts of diammonium glycyrrhizinate, 55 parts of fish collagen peptide, 12 parts of soybean peptide, 15 parts of stem cell exosome, 1.5 parts of SHH protein, 1.5 parts of SMO inhibitor, 0.5 parts of HhSP protein, 6 parts of N-acetyl tyrosine, 0.2 parts of hyaluronic acid, 0.2 parts of melatonin, 0.2 parts of quercetin, 0.2 parts of kaempferol, 0.2 parts of tanshinone, 0.2 parts of biotin and 0.2 parts of vitamin A.

[0026] The preparation method of biotin tripeptide-1 is: biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide are dissolved in tetrahydrofuran, heated to 60℃, and refluxed for 1h to prepare biotin tripeptide-1; the mass ratio of biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide is 5:3:1.

[0027] The fish collagen peptide is extracted by protease for 6h, and then compound protease is added after enzyme inactivation to prepare it by secondary enzymolysis, and the compound protease includes bromelain, neutral protease, alkaline protease and flavor protease, and the mass ratio is 1:2:1:2.

[0028] The enzymolysis conditions in fish collagen peptide are: the temperature is 40℃, the pH is 2.5, the enzymolysis time is 3h, and the enzyme addition amount is 1.5% of the raw material.

[0029] The stem cell exosome is extracted from the adipose tissue near the rat groin and testis.

[0030] A preparation method of an exosome composite preparation with hair growth efficacy, comprising the following steps: (1) N-acetyl tyrosine, hyaluronic acid, melatonin, quercetin, kaempferol, tanshinone, biotin and vitamin A are treated into stem cell exosomes by ultrasonic treatment to obtain exosome-loaded active extract; (2) Biotin tripeptide-1, fish collagen peptide, and soybean peptide are mixed with the exosome-loaded active extract to prepare a loaded active peptide; (3) Ginsenoside Rb1, SHH protein, SMO inhibitor, HhSP protein, and diammonium glycyrrhizinate are mixed with the loaded active peptide to prepare a mixed composite preparation.

[0031] Example 3 An exosome composite preparation with hair growth efficacy, comprising the following components: ginsenoside Rb1 16 parts, biotin tripeptide-1 7 parts, diammonium glycyrrhizinate 10 parts, fish collagen peptide 60 parts, soybean peptide 14 parts, stem cell exosomes 15 parts, SHH protein 2.0 parts, SMO inhibitor 2.0 parts, HhSP protein 0.5 parts, N-acetyl tyrosine 7 parts, hyaluronic acid 0.3 parts, melatonin 0.3 parts, quercetin 0.3 parts, kaempferol 0.3 parts, tanshinone 0.3 parts, biotin 0.3 parts, and vitamin A 0.3 parts.

[0032] The preparation method of biotin tripeptide-1 is as follows: biotin, N-hydroxysuccinimide, and 1,3-dicyclohexyl carbodiimide are dissolved in tetrahydrofuran, heated to 70℃, and refluxed for 2h to obtain biotin tripeptide-1; the mass ratio of biotin, N-hydroxysuccinimide, and 1,3-dicyclohexyl carbodiimide is 7:2:1.

[0033] The fish collagen peptide is extracted by protease for 5h, and then compound protease is added after enzyme inactivation to prepare it by secondary enzymolysis; the compound protease includes bromelain, neutral protease, alkaline protease, and flavor protease, and the mass ratio is 2:2:1:3.

[0034] The enzymolysis conditions of fish collagen peptide are as follows: the temperature is 42℃, the pH is 2.8, the enzymolysis time is 5h, and the enzyme addition amount is 2.5% of the raw material.

[0035] The stem cell exosomes are extracted from the adipose tissue near the rat inguinal and testis.

[0036] A preparation method of an exosome composite preparation with hair growth efficacy, comprising the following steps: (1) N-acetyl tyrosine, hyaluronic acid, melatonin, quercetin, kaempferol, tanshinone, biotin and vitamin A are treated into stem cell exosomes by ultrasonic treatment to obtain exosome-loaded active extract; (2) Biotin tripeptide-1, fish collagen peptide, soybean peptide are mixed with the exosome-loaded active extract to prepare the active peptide-loaded preparation. (3) Ginsenoside Rb1, SHH protein, SMO inhibitor, HhSP protein and diammonium glycyrrhizinate are mixed with the active peptide-loaded preparation to prepare the mixed composite preparation.

[0037] Example 4 An exosome composite preparation with hair growth efficacy comprises the following components: 18 parts of ginsenoside Rb1, 6 parts of biotin tripeptide-1, 10 parts of diammonium glycyrrhizinate, 64 parts of fish collagen peptide, 14 parts of soybean peptide, 15 parts of stem cell exosome, 3.2 parts of SHH protein, 1.8 parts of SMO inhibitor, 1.0 part of HhSP protein, 7 parts of N-acetyl tyrosine, 0.5 part of hyaluronic acid, 0.5 part of melatonin, 0.5 part of quercetin, 0.5 part of kaempferol, 0.5 part of tanshinone, 0.5 part of biotin and 0.5 part of vitamin A.

[0038] The preparation method of biotin tripeptide-1 is as follows: biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide are dissolved in tetrahydrofuran, heated to 60℃, and refluxed for 1h to obtain biotin tripeptide-1; the mass ratio of biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide is 5:3:1.

[0039] The fish collagen peptide is prepared by extracting with protease for 4h and then adding compound protease for secondary enzymolysis after enzyme inactivation; the compound protease includes bromelain, neutral protease, alkaline protease and flavor protease, and the mass ratio is 1:2:1:2.

[0040] The enzymolysis conditions of fish collagen peptide are as follows: the temperature is 45℃, the pH is 3.8, the enzymolysis time is 6h, and the enzyme addition amount is 4.2% of the raw material.

[0041] The stem cell exosome is extracted from the adipose tissue near the rat inguinal and testis.

[0042] A preparation method of an exosome composite preparation with hair growth efficacy comprises the following steps: (1) N-acetyl tyrosine, hyaluronic acid, melatonin, quercetin, kaempferol, tanshinone, biotin and vitamin A are treated into stem cell exosomes by ultrasonic treatment to obtain exosome-loaded active extract; (2) mixing the biotin tripeptide-1, fish collagen peptide, and soybean peptide with the exosome-loaded active extract to obtain a loaded active peptide; (3) mixing the ginsenoside Rb1, SHH protein, SMO inhibitor, HhSP protein, and diammonium glycyrrhizinate with the loaded active peptide to obtain a mixed composite preparation.

[0043] Example 6 An exosome composite preparation with hair growth efficacy comprises the following components: 19 parts of ginsenoside Rb1, 9 parts of biotin tripeptide-1, 11 parts of diammonium glycyrrhizinate, 58 parts of fish collagen peptide, 15 parts of soybean peptide, 21 parts of stem cell exosome, 3.8 parts of SHH protein, 2.1 parts of SMO inhibitor, 1.5 parts of HhSP protein, 8 parts of N-acetyl tyrosine, 0.8 parts of hyaluronic acid, 0.6 parts of melatonin, 0.6 parts of quercetin, 0.6 parts of kaempferol, 0.6 parts of tanshinone, 0.6 parts of biotin, and 0.6 parts of vitamin A.

[0044] The preparation method of the biotin tripeptide-1 is as follows: dissolving biotin, N-hydroxysuccinimide, and 1,3-dicyclohexyl carbodiimide in tetrahydrofuran, heating to 70℃, and refluxing for 2h to obtain the biotin tripeptide-1; the mass ratio of biotin, N-hydroxysuccinimide, and 1,3-dicyclohexyl carbodiimide is 7:3:1.

[0045] The fish collagen peptide is prepared by extracting for 6h with a protease and then adding a compound protease for secondary enzymolysis after enzyme inactivation; the compound protease includes bromelain, neutral protease, alkaline protease, and flavor protease, and the mass ratio is 2:1:1:3.

[0046] The enzymolysis conditions of the fish collagen peptide are as follows: the temperature is 50℃, the pH is 7.0, the enzymolysis time is 10h, and the enzyme addition amount is 5.5% of the raw material.

[0047] The stem cell exosome is extracted from the adipose tissue near the rat inguinal and testicular.

[0048] A preparation method of an exosome composite preparation with hair growth efficacy comprises the following steps: (1) treating N-acetyl tyrosine, hyaluronic acid, melatonin, quercetin, kaempferol, tanshinone, biotin, and vitamin A into stem cell exosomes by ultrasonic treatment to obtain an exosome-loaded active extract; (2) mixing the biotin tripeptide-1, fish collagen peptide, and soybean peptide with the exosome-loaded active extract to obtain a loaded active peptide; (3) mixing the ginsenoside Rb1, SHH protein, SMO inhibitor, HhSP protein, and diammonium glycyrrhizinate with the loaded active peptide to obtain a mixed composite preparation.

[0049] Example 7 An exosome composite preparation with hair growth efficacy, comprising the following components: ginsenoside Rb1 20 parts, biotin tripeptide-1 11 parts, diammonium glycyrrhizinate 14 parts, fish collagen peptide 60 parts, soybean peptide 18 parts, stem cell exosome 24 parts, SHH protein 4.0 parts, SMO inhibitor 2.2 parts, HhSP protein 1.2 parts, N-acetyl tyrosine 8 parts, hyaluronic acid 0.8 parts, melatonin 0.8 parts, quercetin 0.8 parts, kaempferol 0.8 parts, tanshinone 0.8 parts, biotin 0.8 parts, and vitamin A 0.8 parts.

[0050] The preparation method of biotin tripeptide-1 is as follows: biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide are dissolved in tetrahydrofuran, heated to 80℃, and refluxed for 2h to obtain biotin tripeptide-1; the mass ratio of biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide is 8:2:1.

[0051] The fish collagen peptide is prepared by extracting with protease for 5h and then adding complex protease for secondary enzymolysis after enzyme inactivation, and the complex protease includes bromelain, neutral protease, alkaline protease and flavor protease, and the mass ratio is 2:1:1:2.

[0052] The enzymolysis conditions of fish collagen peptide are as follows: the temperature is 50℃, the pH is 7.5, the enzymolysis time is 9h, and the enzyme addition amount is 6.2% of the raw material.

[0053] The stem cell exosome is extracted from the adipose tissue near the rat inguinal and testis.

[0054] A preparation method of an exosome composite preparation with hair growth efficacy, comprising the following steps: (1) N-acetyl tyrosine, hyaluronic acid, melatonin, quercetin, kaempferol, tanshinone, biotin and vitamin A are treated by ultrasonic treatment into stem cell exosome to obtain exosome-loaded active extract; (2) Biotin tripeptide-1, fish collagen peptide, soybean peptide and exosome-loaded active extract are mixed to prepare a loaded active peptide; (3) Ginsenoside Rb1, SHH protein, SMO inhibitor, HhSP protein and diammonium glycyrrhizinate are mixed to modify the loaded active peptide to prepare a mixed composite preparation.

[0055] Example 8 An exosome composite preparation with hair growth efficacy, comprising the following components: ginsenoside Rb1 22 parts, biotin tripeptide-1 12 parts, diammonium glycyrrhizinate 15 parts, fish collagen peptide 70 parts, soybean peptide 20 parts, stem cell exosome 25 parts, SHH protein 4.5 parts, SMO inhibitor 2.5 parts, HhSP protein 0.5 parts, N-acetyl tyrosine 10 parts, hyaluronic acid 2 parts, melatonin 1 part, quercetin 1 part, kaempferol 1 part, tanshinone 1 part, biotin 1 part, and vitamin A 1 part.

[0056] The preparation method of biotin tripeptide-1 is as follows: biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide are dissolved in tetrahydrofuran, heated to 90℃, and refluxed for 3h to obtain biotin tripeptide-1; the mass ratio of biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide is 8:3:1.

[0057] The fish collagen peptide is prepared by extracting with protease for 9h and then adding complex protease for secondary enzymolysis after enzyme inactivation, and the complex protease includes bromelain, neutral protease, alkaline protease and flavor protease with a mass ratio of 2:2:1:3.

[0058] The enzymolysis conditions of the fish collagen peptide are as follows: the temperature is 60℃, the pH is 8.5, the enzymolysis time is 12h, and the enzyme addition amount is 8% of the raw material.

[0059] The stem cell exosome is extracted from the adipose tissue near the rat inguinal and testis.

[0060] A preparation method of an exosome composite preparation with hair growth efficacy, comprising the following steps: (1) N-acetyl tyrosine, hyaluronic acid, melatonin, quercetin, kaempferol, tanshinone, biotin and vitamin A are treated by ultrasonic treatment into stem cell exosome to obtain exosome-loaded active extract; (2) Biotin tripeptide-1, fish collagen peptide, soybean peptide and the exosome-loaded active extract are mixed to prepare a loaded active peptide; (3) Ginsenoside Rb1, SHH protein, SMO inhibitor, HhSP protein and diammonium glycyrrhizinate are mixed to modify the loaded active peptide to prepare a mixed composite preparation.

[0061] Comparative Example 1 The preparation method of Example 1 is different in that ginsenoside Rb1 is not added.

[0062] Comparative Example 2 The preparation method of Example 1 is different in that SHH protein is not added.

[0063] Comparative Example 3 The preparation method of Example 1 is different in that no SMO inhibitor is added.

[0064] Comparative Example 4 The preparation method of Example 1 is different in that no stem cell exosome is added.

[0065] Test results: 80 volunteers (40 males and 40 females) were recruited, of which 30 volunteers had dry hair, 32 volunteers had oily hair, and 18 volunteers had damaged hair. It was ensured that the volunteers did not use other special products (such as hair dyes and hair relaxers) that could affect the state of the hair within a certain period of time (such as 2 weeks) before the experiment.

[0066] The volunteers were randomly divided into a test group and a control group. The volunteers tried to keep other hair care habits unchanged during the experiment. The test group used the hair loss prevention conditioner three times a week, and the control group did not use the conditioner.

[0067] Before and after the experiment, the hair breakage strength, frizz level, and moisture retention rate of all volunteers were measured, and the collected data was statistically analyzed after 8 weeks.

[0068] Human trials showed that continuous use of the exosome complex formulation containing 5% w / v (30 mg / mL) with hair growth efficacy for 8 weeks increased the hair breakage strength by 28%, reduced the frizz level by 40%, and increased the moisture retention rate by 23%.

[0069] Animal experiments: Forty male B6CBAF1 / J mice were used to model by subcutaneous injection of testosterone propionate on the back of the mice. After successful modeling, they were randomly divided into 4 groups, 10 in each group. Examples 1, 2 and 3 were given the corresponding mixed exosome complex formulation with hair growth efficacy, 2 times a day, massaging for 5 minutes; The blank group was not given any medication.

[0070] After 4 weeks of experiment, the growth of hair of the experimental animals in the 4 groups was observed, and the depilation area on the back of the mice was taken, with an area of 1x1 cm and a depth of fascia. The effect on the number of mouse hair follicles was explored, and the average value was taken. The results are shown in Table 1.

[0071] Table 1 The above is only an exemplary specific embodiment of the present application, but the protection scope of the present application is not limited thereto. Any changes or replacements within the technical range disclosed by the present application can be easily thought of by those skilled in the art, and should be covered within the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. An exosome complex preparation having hair growth efficacy, characterized by, The substance comprises the following components: 12-22 parts of ginsenoside Rb1, 5-12 parts of biotin tripeptide-1, 8-15 parts of diammonium glycyrrhizinate, 50-70 parts of fish collagen peptide, 10-20 parts of soybean peptide, 12-25 parts of stem cell exosome, 0.5-4.5 parts of SHH protein, 0.5-2.5 parts of SMO inhibitor, 0.5-1.5 parts of HhSP protein, 5-10 parts of N-acetyl tyrosine, 0.1-2 parts of hyaluronic acid, 0.1-1 part of melatonin, 0.1-1 part of quercetin, 0.1-1 part of kaempferol, 0.1-1 part of tanshinone, 0.1-1 part of biotin and 0.1-1 part of vitamin A.

2. The exosome complex formulation having hair growth efficacy according to claim 1, wherein, The preparation method of the biotin tripeptide-1 is as follows: dissolving biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide in tetrahydrofuran, heating to 60-90 DEG C, refluxing for 1-3 hours to obtain the biotin tripeptide-1; the mass ratio of the biotin, N-hydroxysuccinimide and 1,3-dicyclohexyl carbodiimide is 5-8:2-3:

1. 3.The exosome complex formulation having hair growth efficacy according to claim 1, characterized in that, The fish collagen peptide is prepared by extracting with protease for 4-9 hours, adding complex protease after enzyme inactivation for secondary enzymolysis, and the complex protease comprises at least one of bromelain, neutral protease, alkaline protease and flavor protease.

4. The exosome complex formulation having hair growth efficacy according to claim 3, characterized by, The enzymolysis conditions of the fish collagen peptide are as follows: the temperature is 37-60 DEG C, the pH is 1.5-8.5, the enzymolysis time is 1-12 hours, and the enzyme addition amount is 0.5%-8% of the raw material. 5.The exosome complex formulation having hair growth efficacy according to claim 1, characterized in that, The stem cell exosome is ipsc-derived, the total yield of the stem cell exosome is 2.5 x 10 4 ; the HhSP protein is a hedgehog pathway protein.

6. The method of claim 1-5, wherein the exosome complex formulation having hair growth efficacy is prepared by the following steps of: (a) mixing the exosome complex formulation having hair growth efficacy with a pharmaceutically acceptable carrier; (b) adding a pharmaceutically acceptable additive to the mixture; and (c) sterilizing the mixture. The method comprises the following steps: (1) treating N-acetyl tyrosine, hyaluronic acid, melatonin, quercetin, kaempferol, tanshinone, biotin and vitamin A by ultrasonic treatment into stem cell exosome to obtain exosome-loaded active extract; (2) mixing biotin tripeptide-1, fish collagen peptide, soybean peptide and the exosome-loaded active extract to prepare loaded active peptide; (3) mixing ginsenoside Rb1, SHH protein, SMO inhibitor, HhSP protein and diammonium glycyrrhizinate to modify the loaded active peptide to prepare a mixed composite preparation.

7. Application of the exosome composite preparation with hair growth efficacy according to claim 1 in the preparation of a product for preventing hair loss, promoting hair growth and hair growth.

8. Use according to claim 7, characterized in that, The product for preventing hair loss, promoting hair growth and hair growth is a drug or a care product; and / or The dosage form of the product for preventing hair loss, promoting hair growth and hair growth comprises any one of the following: a coating agent, a spray, a lotion, a paste, a rubbing agent and a gel.

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